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PubMed1997

Dielectrophoretic detection of changes in erythrocyte membranes following malarial infection.

The dielectric properties of normal erythrocytes were compared to those of cells infected with the malarial parasite Plasmodium falciparum. Normal cells provided stable electrorotation spectra which, when analyzed by a single-shelled oblate spheroid dielectric model, gave a specific capacitance value of 12 +/- 1.2 mF/m2 for the plasma membrane, a cytoplasmic permittivity of 57 +/- 5.4 and a cytoplasmic conductivity of 0.52 +/- 0.05 S/m. By contrast, parasitized cells exhibited electrorotation spectra with a time-dependency that suggested significant net ion outflux via the plasma membrane and it was not possible to derive reliable cell parameter values in this case. To overcome this problem, cell membrane dielectric properties were instead determined from dielectrophoretic crossover frequency measurements made as a function of the cell suspending medium conductivity. The crossover frequency for normal cells depended linearly on the suspension conductivity above 20 mS/m and analysis according to the single-shelled oblate spheroid dielectric model yielded values of 11.8 mF/m2 and 271 S/m2, respectively, for the specific capacitance and conductance of the plasma membrane. Unexpectedly, the crossover frequency characteristics of parasitized cells at high suspending medium conductivities were non-linear. This effect was analyzed in terms of possible dependencies of the cell membrane capacitance, conductance or shape on the suspension medium conductivity, and we concluded that variations in the membrane conductance were most likely responsible for the observed non-linearity. According to this model, parasitized cells had a specific membrane capacitance of 9 +/- 2 mF/m2 and a specific membrane conductance of 1130 S/m2 that increased with increasing cell suspending medium conductivity. Such conductivity changes in parasitized cells are discussed in terms of previously observed parasite-associated membrane pores. Finally, we conclude that the large differences between the dielectrophoretic crossover characteristics of normal and parasitized cells should allow straightforward sorting of these cell types by dielectrophoretic methods.

Animals↗

PubMed2005

Vibrations and diverging length scales near the unjamming transition.

We numerically study the vibrations of jammed packings of particles interacting with finite-range, repulsive potentials at zero temperature. As the packing fraction phi is lowered towards the onset of unjamming at phi(c), the density of vibrational states approaches a nonzero value in the limit of zero frequency. For phi >phi(c), there is a crossover frequency, omega* below which the density of states drops towards zero. This crossover frequency obeys power-law scaling with phi-phi(c). Characteristic length scales, determined from the dominant wave vector contributing to the eigenmode at omega*, diverge as power laws at the unjamming transition.

Journal Article↗

PubMed1986

The pattern of pairing that is effective for crossing over in complex B-A chromosome rearrangements in maize. III. Possible evidence for pairing centers.

The study of the mechanism of meiotic homolog pairing, approached by comparing chiasma frequencies in rearranged segments that differ in relative length and intrachromosomal location, is substantially extended here. For the first time, two kinds of evidence were found that centers specialized for alignment pairing may exist in maize chromosomes: for two segments, higher than average crossover frequency per unit length was maintained when these were located in several different chromosomal positions with respect to centromere and telomere, and in fact apart from their own normal centromeres and telomeres. High crossover frequencies in these segments regardless of position are considered to reflect innate capacity for alignment pairing due to relatively strong pairing center content. For a short rearranged segment, chiasma frequency was drastically reduced, and evidence suggests that all of the chiasmata found there depended upon juxtaposition made possible by the completion of the zip-up pairing process in the other arms of the translocation configuration. This short segment is thought to be essentially devoid of pairing center content. It seems possible that crossover frequency depression in short rearranged segments may usually not be due, as commonly supposed, to mechanical difficulties inherent in formation of contorted configurations, but rather to absence of pairing centers within them and the relative rarity (compared to the normal sequence situation) of enabling zip-up pairing. Evidence also indicates that pairing which leads to crossing over must frequently occur between internal translocated segments and their normal sequence counterparts in a way which cannot be dependent upon zipping-up of two-by-two pairing initiated at or near telomeres. Pairing centers in maize are probably numerous and widely dispersed, since coarse direct proportionality is found when chiasma frequency is compared for an array of segment lengths.

Centromere↗

PubMed1992

A linkage map of mouse chromosome 1 using an interspecific cross segregating for the gld autoimmunity mutation.

An interspecific backcross was used to define a high resolution linkage map of mouse Chromosome (Chr) 1 and to analyze the segregation of the generalized lymphoproliferative disease (gld) mutation. Mice homozygous for gld have multiple features of autoimmune disease. Analysis of up to 428 progeny from the backcross [(C3H/HeJ-gld x Mus spretus)F1 x C3H/HeJ-gld] established a map that spans 77.6 cM and includes 56 markers distributed over 34 ordered genetic loci. The gld mutation was mapped to a less than 1 cM segment on distal mouse Chr 1 using 357 gld phenotype-positive backcross mice. A second backcross, between the laboratory strains C57BL/6J and SWR/J, was examined to compare recombination frequency between selected markers on mouse Chr 1. Significant differences in crossover frequency were demonstrated between the interspecific backcross and the inbred laboratory cross for the entire interval studied. Sex difference in meiotic crossover frequency was also significant in the laboratory mouse cross. Two linkage groups known to be conserved between segments of mouse Chr 1 and the long arm of human Chrs 1 and 2 where further defined and a new conserved linkage group was identified that includes markers of distal mouse Chr 1 and human Chr 1, bands q32 to q42.

Animals↗

PubMedDate not supplied

Acoustic analysis of speech through a hearing aid: consonant-vowel ratio effects with two-channel compression amplification.

In this study, the consonant-vowel ratio (CVR) changes associated with varying the compression ratio and crossover frequency of two-channel syllabic compression amplification were examined. Consonant-vowel syllables were recorded at 60 and 75 dB SPL input levels to the hearing aid under 13 different amplification conditions: 12 compression conditions and a condition that used linear amplification in both channels. Syllables consisted of voiceless stops, fricatives, and affricates combined with the vowels /a/, /i/, and /u/. Acoustic analysis of the processed syllables indicated that the CVR was generally increased with compression, compared to linear amplification, and that the effects were greatest for amplification with compression in the high-frequency channel. In addition, higher CVRs were obtained when the crossover frequency was raised. Compression in the low-frequency channel had the least effect on CVR. As previous research has indicated that CVR may serve as a cue to the perception of some consonant sounds by people with hearing impairment, the results suggest the need for caution with the application of high-frequency channel compression until the perceptual effects are fully investigated.

Hearing Aids↗

PubMed1987

A frequency importance function for continuous discourse.

Normal hearing subjects estimated the intelligibility of continuous discourse (CD) passages spoken by three talkers (two male and one female) under 135 conditions of filtering and signal-to-noise ratio. The relationship between the intelligibility of CD and the articulation index (the transfer function) was different from any found in ANSI S3.5-1969. Also, the lower frequencies were found to be relatively more important for the intelligibility of CD than for identification of nonsense syllables and other types of speech for which data are available except for synthetic sentences [Speaks, J. Speech Hear. Res. 10, 289-298 (1967)]. The frequency which divides the auditory spectrum into two equally important halves (the crossover frequency) was found to be about 0.5 oct lower for the CD used in this study than the crossover frequency for male talkers of nonsense syllables found in ANSI S3.5-1969 and about 0.7 oct lower than the one for combined male and female talkers of nonsense syllables reported by French and Steinberg [J. Acoust. Soc. Am. 19, 90-119 (1947)].

Adolescent↗

PubMed2000

Recombination in an isochromosome preferentially occurs between cis isochromatids.

An isochromosome has identical arms attached to the same centromere. At the pachytene stage of meiosis, it has four isochromatids and recombination can occur either between cis isochromatids (attached to the same half-centromere) or trans isochromatids (attached to different half-centromeres). Normally such recombination cannot be detected because all four chromatids are homogenetic (arose from misdivision of a centromere to which genetically identical sister chromatids were attached). We isolated an isochromosome of wheat that is heterogenetic for the distal 64% of the long arm. The heterogenetic isochromosome was recovered from the progeny of a cross between Triticum aestivum cv. Chinese Spring containing an isochromosome for the long arm of chromosome 5B (i5BL) and a disomic substitution line of Triticum turgidum ssp. dicoccoides chromosome 5B in Chinese Spring wheat. New recombinants were produced when the two arms of i5BLrec paired at metaphase I of meiosis. Only trans isochromatid exchanges led to some homozygous loci in i5BLrec, whereas exchanges between cis isochromatids resulted in heterozygosity at all loci similar to the parental type. There was an average frequency of 0.87 chiasmata per pollen mother cell for the heterogenetic i5BL, which will result in 0.44 cis and 0.44 trans isochromatid exchanges, assuming that both are occurring at the same frequency. The average crossover frequency based on recombination between trans isochromatid exchange detected by restriction fragment length polymorphism analysis in 98 plants was 0.29. This observed value is significantly lower (P<0.01) than the value of 0.44 as expected from chiasmata counts. Our study provides the first experimental evidence that crossovers preferentially occur between cis isochromatids rather than trans isochromatids.

Chromatids↗

PubMed2006

DM2 CCTGCAGG repeats are crossover hotspots that are more prone to expansions than the DM1 CTGCAG repeats in Escherichia coli.

Myotonic dystrophy type 2 (DM2) is caused by the extreme expansion of the repeating tetranucleotide CCTG*CAGG sequence from <30 repeats in normal individuals to approximately 11,000 for the full mutation in certain patients. This repeat is in intron 1 of the zinc finger protein 9 gene on chromosome 3q21. Since prior work demonstrated that CTG*CAG and GAA*TTC triplet repeats (responsible for DM1 and Friedreich's ataxia, respectively) can expand by genetic recombination, we investigated the capacity of the DM2 tetranucleotide repeats to also expand during this process. Both gene conversion and unequal crossing over are attractive mechanisms to effect these very large expansions. (CCTG*CAGG)n (where n=30, 75, 114 or 160) repeats showed high recombination crossover frequencies (up to 27-fold higher than the non-repeating control) in an intramolecular plasmid system in Escherichia coli. Furthermore, a distinct orientation effect was observed where orientation II (CAGG on the leading strand template) was more prone to recombine. Expansions of up to double the length of the tetranucleotide repeats were found. Also, the repeating tetranucleotide sequence was more prone to expansions (to give lengths longer than a single repeating tract) than deletions as observed for the CTG*CAG and GAA*TTC repeats. We determined that the DM2 tetranucleotide repeats showed a lower thermodynamic stability when compared to the DM1 trinucleotide repeats, which could make them better targets for DNA repair events, thus explaining their expansion-prone behavior. Genetic studies in SOS-repair mutants revealed high frequencies of recombination crossovers although the SOS-response itself was not induced. Thus, the genetic instabilities of the CCTG*CAGG repeats may be mediated by a recombination-repair mechanism that is influenced by DNA structure.

Chromosome Mapping↗

PubMed1979

Programmable hearing aid with multichannel compression.

A portable programmable hearing aid with multichannel compression was constructed with three channels with two independently variable crossover frequencies. The compression knee-point and attenuation in each channel and the two crossover frequencies can be programmed from a separate programming unit. Speech discrimination for seven subjects with sensorineural hearing loss was evaluated, comparing each subject individually in three different cases--equalization, multichannel compression and the subject's own conventional hearing aid. Each subject wore each aid for about a week in his/her own environment and was then tested. The following speech discrimination tests were used: monosyllabic phonetically balanced words with and without background noise, nonsense syllables in quiet, and a constant test in quiet and in noise. The study showed that some patients with sensorineural hearing loss accompanied by recruitment can double their speech discrimination by using a hearing aid with multichannel compression.

Hearing Aids↗

PubMed1972

The temporal sequence of synaptic initiation, crossing over and synaptic completion.

Questions are raised as to the validity of arguments that crossover positions have been demonstrated to be normally established only during pachytene (after synapsis is maximal). An alternative and testable hypothesis is that crossover commitment can occur at events of synaptic initiation.-Measurements are presented of extents of pachytene synapsis and failure in and around a region of maize chromosome heterozygous for a short paracentric inversion, and these are compared to conjectured expectations from observations of crossover frequencies within the inversion. Various hypotheses consistent with the results are considered. It is pointed out that the hypothesis that increases in crossover frequency in the synapsed region of the inversion are compensatory to crossover inhibitions elsewhere requires complex assumptions: that the adjustment must take place among, not within cells and that the enhancement is preferentially expressed within the inversion instead of elsewhere in the genome. The hypothesis that the fixing and squashing procedure forces apart non-crossover regions previously synapsed but lacking a crossover also requires complex assumptions. The simplest hypothesis proposes that crossover commitment may determine synaptic expression. A role of the synaptonemal complex in the establishment of crossover sites is questioned or minimized.-Evidence is also presented with respect to conceivable function of the telomere in synaptic initiation. Restrictions on such a function, if it exists, seem to be required to account for the observations.

Chromosome Aberrations↗

PubMed1997

Characterization of susceptible chiasma configurations that increase the risk for maternal nondisjunction of chromosome 21.

Recent studies of trisomy 21 have shown that altered levels of recombination are associated with maternal non-disjunction occurring at both meiosis I (MI) and meiosis II (MII). To comprehend better the association of recombination with nondisjunction, an understanding of the pattern of meiotic exchange, i.e. the exchange of genetic material at the four-strand stage during prophase, is required. We examined this underlying exchange pattern to determine if specific meiotic configurations are associated with a higher risk of non-disjunction than others. We examined the crossover frequencies of chromosome 21 for three populations: (i) normal female meiotic events; (ii) meiotic events leading to MI non-disjunction; and (iii) those leading to MII non-disjunction. From these crossover frequencies, we estimated the array of meiotic tetrads that produced the observed crossovers. Using this approach, we found that nearly one-half of MI errors were estimated to be achiasmate. The majority of the remaining MI bivalents had exchanges that clustered at the telomere. In contrast, exchanges occurring among MII cases clustered at the pericentromeric region of the chromosome. Unlike the single exchange distributions, double exchanges from the non-disjoined populations seemed to approximate the distribution in the normal population. These data suggest that the location of certain exchanges makes a tetrad susceptible to non-disjunction. Specifically, this susceptibility is associated with the distance between the centromere and closest exchange. This result challenges the widely held concept that events occurring at MII are largely independent of events occurring at MI, and suggests that all non-disjunction events may be initiated during MI and simply resolved at either of the two meiotic stages.

Chromosomes, Human, Pair 21↗

PubMed2006

Polymorphism in the activity of human crossover hotspots independent of local DNA sequence variation.

Meiotic crossovers in the human genome cluster into highly localized hotspots identifiable indirectly from patterns of DNA diversity and directly by high-resolution sperm typing. Little is known about factors that control hotspot activity and the apparently rapid turnover of hotspots during recent evolution. Clues can, however, be gained by characterizing variation in sperm crossover activity between men. Previous studies have identified single nucleotide polymorphisms within hotspots that appear to suppress crossover activity and which may be involved in hotspot attenuation/extinction. We now analyse a closely spaced pair of hotspots (MSTM1a, MSTM1b) on chromosome 1q42.3, the former being a candidate for a young hotspot that has failed to leave a significant mark on haplotype diversity. Extensive surveys of different men revealed substantial polymorphism in sperm crossover frequencies at both hotspots, but with very different patterns of variation. Hotspot MSTM1b was active in all men tested but with widely differing crossover frequencies. In contrast, MSTM1a was active in only a few men and appeared to be recombinationally inert in the remainder, providing the first example of presence/absence polymorphism of a human hotspot. Haplotype analysis around both hotspots identified active and suppressed men sharing identical haplotypes, establishing that these major variations in the presence/absence of a hotspot and in quantitative activity are not caused by local DNA sequence variation. These findings suggest a role for distal regulators or epigenetic factors in hotspot activity and provide the first direct evidence for the rapid evolution of recombination hotspots in humans.

Chromosome Mapping↗

PubMed2002

Male mouse recombination maps for each autosome identified by chromosome painting.

Linkage maps constructed from genetic analysis of gene order and crossover frequency provide few clues to the basis of genomewide distribution of meiotic recombination, such as chromosome structure, that influences meiotic recombination. To bridge this gap, we have generated the first cytological recombination map that identifies individual autosomes in the male mouse. We prepared meiotic chromosome (synaptonemal complex [SC]) spreads from 110 mouse spermatocytes, identified each autosome by multicolor fluorescence in situ hybridization of chromosome-specific DNA libraries, and mapped >2,000 sites of recombination along individual autosomes, using immunolocalization of MLH1, a mismatch repair protein that marks crossover sites. We show that SC length is strongly correlated with crossover frequency and distribution. Although the length of most SCs corresponds to that predicted from their mitotic chromosome length rank, several SCs are longer or shorter than expected, with corresponding increases and decreases in MLH1 frequency. Although all bivalents share certain general recombination features, such as few crossovers near the centromeres and a high rate of distal recombination, individual bivalents have unique patterns of crossover distribution along their length. In addition to SC length, other, as-yet-unidentified, factors influence crossover distribution leading to hot regions on individual chromosomes, with recombination frequencies as much as six times higher than average, as well as cold spots with no recombination. By reprobing the SC spreads with genetically mapped BACs, we demonstrate a robust strategy for integrating genetic linkage and physical contig maps with mitotic and meiotic chromosome structure.

Adaptor Proteins, Signal Transducing↗

PubMed1999

The Saccharomyces cerevisiae MER3 gene, encoding a novel helicase-like protein, is required for crossover control in meiosis.

The MER3 gene is identified as a novel meiosis-specific gene, whose transcript is spliced in an MRE2/MER1-dependent manner. The predicted Mer3 protein contains the seven motifs characteristic of the DExH-box type of helicases as well as a putative zinc finger. Double strand breaks (DSBs), the initial changes of DNA in meiotic recombination, do not disappear completely and are hyperresected late in mer3 meiosis, indicating that MER3 is required for the transition of DSBs to later intermediates. A mer3 mutation reduces crossover frequencies, and the remaining crossovers show random distribution along a chromosome, resulting in a high incidence of non-disjunction of homologous chromosomes at the first meiotic division. MER3 appears to be very important for both the DSB transition and crossover control.

Amino Acid Sequence↗

PubMed2003

Frequency characteristics of long-term heart rate variability during constant-routine protocol.

The effects of such behavioral factors as physical activity, food intake, and circadian rhythm on long-term heart rate variability (HRV) in humans remain poorly understood. We therefore studied their effects on HRV using a constant-routine protocol that included simultaneous core body temperature (CBT) correction. Seven healthy subjects completed the constant-routine and daily-routine protocols, during which HRV and CBT were continuously monitored. During the constant routine, subjects were kept awake for 27 h in a semirecumbent posture with minimal physical activity; small isocaloric meals were provided every 2 h. During the daily routine, subjects carried on their lives normally. Data were analyzed using generic spectral analysis based on a fast Fourier transform; coarse-graining spectral analysis was also used to eliminate periodicity due to the regular meals for raw HRV and for the CBT-corrected HRV without circadian and/or low-frequency ultradian components. The results showed that 1) the power spectra of HRV in the constant routine and daily routine had similar power-law scalings at frequencies above approximately 10(-3.5) Hz, while 2) below that crossover frequency, HRV was smaller in the constant routine than in the daily routine, with the difference becoming significant (P < 0.05) at <10(-4) Hz, 3) coarse-graining spectral analysis eliminated diet-induced peaks in generic spectral analysis-based HRV spectra during the constant routine and emphasized the crossover at approximately 10(-3.5) Hz, and 4) CBT correction did not alter the results. Below a frequency of approximately 10(-3.5) Hz (a period >1 h), HRV is strongly influenced by behavioral factors; above that crossover frequency, HRV is behavior independent, possibly reflecting an intrinsic regulatory system.

Adult↗

PubMed2002

[Establishment of gene replacement/disruption system through homologous recombination in Amycolatopsis mediterranei U32].

A gene replacement/disruption system of Amycolatopsis mediterranei U32 was developed based on the established electroporation conditions as well as appropriate selective markers. Through two-step selection, ahbas gene in U32 was replaced by a promoterless alpha-amylase gene constructed on the plasmid pDK110 of E. coli. The first single-crossover and the second double-crossover frequencies were approximately 0.5%-0.7% and 2%, respectively. Denaturation of the plasmid pDK110 increased the integration frequency about 7-10 folds, while electric shock treatment of the single-crossover recombinants increased the frequency of second crossover recombination about 5 folds. Employing denatured DNA fragments containing an apramycin-resistance gene flanked with regions of the respective genes, One-step disruption of rifO and amrA genes of U32 was also achieved with an efficiency of 30-50 transformants per microgram of DNA.

Actinomycetales↗

PubMed2005

Uneven distribution of expressed sequence tag loci on maize pachytene chromosomes.

Examining the relationships among DNA sequence, meiotic recombination, and chromosome structure at a genome-wide scale has been difficult because only a few markers connect genetic linkage maps with physical maps. Here, we have positioned 1195 genetically mapped expressed sequence tag (EST) markers onto the 10 pachytene chromosomes of maize by using a newly developed resource, the RN-cM map. The RN-cM map charts the distribution of crossing over in the form of recombination nodules (RNs) along synaptonemal complexes (SCs, pachytene chromosomes) and allows genetic cM distances to be converted into physical micrometer distances on chromosomes. When this conversion is made, most of the EST markers used in the study are located distally on the chromosomes in euchromatin. ESTs are significantly clustered on chromosomes, even when only euchromatic chromosomal segments are considered. Gene density and recombination rate (as measured by EST and RN frequencies, respectively) are strongly correlated. However, crossover frequencies for telomeric intervals are much higher than was expected from their EST frequencies. For pachytene chromosomes, EST density is about fourfold higher in euchromatin compared with heterochromatin, while DNA density is 1.4 times higher in heterochromatin than in euchromatin. Based on DNA density values and the fraction of pachytene chromosome length that is euchromatic, we estimate that approximately 1500 Mbp of the maize genome is in euchromatin. This overview of the organization of the maize genome will be useful in examining genome and chromosome evolution in plants.

Chromosome Mapping↗

PubMed1985

Temporal tuning and nonlinearity of intraretinal pathways in turtle: effects of temperature, stimulus intensity, and size.

Flash responses, amplitude and phase transfer functions, and nonlinearities were measured in turtle retina for pathways with photoreceptor inputs and outputs from horizontal (HC), hyperpolarizing bipolar (HBC), sustained amacrine (AC), and on-off ganglion (GC) cells. Flash responses slowed and attenuated in all cells as temperature decreased. White-noise transfer properties of sustained-type cells (HC, HBC, AC) were of low- or bandpass type; high-frequency cut-off (fc) and phase crossover frequency decreased with temperature. fc increased as spot diameter was increased. Nonlinearity of these sustained-response pathways (distortion product frequencies in response to a sum-of-sinusoids input probe) increases with intensity and may depend on amplitude saturation limiting. On/off GC synaptic and spike activity increased as spot diameter decreased and intensity increased. Amplitude transfer functions had a low-frequency peak (PSP activity) and monotonically decreasing amplitude vs. frequency shape (spikes and transient PSP activity). Nonlinearity increased with stimulus intensity; it was maximal with 1 mm spot size, less with smaller (500 micron) and larger (5 mm) spots. It may depend on the functional equivalent of full-wave rectification (on-off response).

Animals↗