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Comparison of vancomycin- and teicoplanin-induced histamine release and "red man syndrome".

Twelve healthy adult males participated in a double-blind, randomized, two-way crossover study to determine histamine release and the frequency and severity of "red man syndrome" (RMS) following intravenous administration of vancomycin (15 mg/kg of body weight over 60 min) and teicoplanin (15 mg/kg over 30 min). Concentrations of vancomycin and teicoplanin in serum and concentrations of histamine in plasma were measured at baseline and during and after each infusion. Erythema and pruritus were classified a priori as mild, moderate, or severe. The extent of erythema was determined by the use of a burn chart, and pruritus was assessed by the subject with a rank scale. Global severity of RMS was determined by summation of the individual scores for pruritus and erythema. Baseline areas under the concentration-time curve for histamine were not significantly different for the vancomycin and teicoplanin treatments. Vancomycin caused RMS in 11 of 12 subjects (9 severe and 2 moderate cases) and was associated with a significant increase in plasma histamine (46.7 +/- 31.3 ng.min/ml, P less than 0.05). In contrast, teicoplanin did not cause RMS or elicit significant histamine release (8.7 +/- 13.2 ng.min/ml). Peak concentrations of vancomycin and teicoplanin in serum were 58.8 +/- 8.4 and 148.0 +/- 31.8 micrograms/ml, respectively (P less than 0.05). Assuming equal efficacy, these data suggest that teicoplanin may be a safe alternative agent in subjects experiencing severe RMS due to vancomycin; however, further studies in the clinical setting are needed.

Adult↗

GENE TRANSFER BY F' STRAINS OF ESCHERICHIA COLI. IV. EFFECT OF A CHROMOSOMAL DELETION ON CHROMOSOME TRANSFER.

Pittard, James (Yale University, New Haven, Conn.), and T. Ramakrishnan. Gene transfer by F' strains of Escherichia coli. IV. The effect of a chromosomal deletion on chromosome transfer. J. Bacteriol. 88:367-373. 1964.-Evidence is presented that in an F' strain of Escherichia coli. AB1206, the chromosomal region corresponding to the merogenote of F(14) is deleted. AB1206 differs from all other F' strains in its stability and in its relative inability to transfer chromosomal markers under conditions in which transfer of the merogenote occurs at a frequency of 100%. Its stability can be accounted for by the fact that the loss of the F merogenote, which contains approximately 10% of the genetic information of the cell, would be a lethal event. The failure of AB1206 to transfer chromosomal markers, except at very low frequencies, can be explained in terms of the crossover model proposed to explain chromosome transfer by F' strains, since the homologous chromosomal region is missing in this strain. The conclusion that the chromosomal region is deleted is strengthened by the finding that the different enzymes coded for by the genes on F(14) are present in AB1206 at the level found in haploid strains, but have double this activity in homozygous diploids carrying F(14).

Chromosomes↗

Gene replacement with one-sided homologous recombination.

Homologous recombination is now routinely used in mammalian cells to replace endogenous chromosomal sequences with transferred DNA. Vectors for this purpose are traditionally constructed so that the replacement segment is flanked on both sides by DNA sequences which are identical to sequences in the chromosomal target gene. To test the importance of bilateral regions of homology, we measured recombination between transferred and chromosomal immunoglobulin genes when the transferred segment was homologous to the chromosomal gene only on the 3' side. In each of the four recombinants analyzed, the 5' junction was unique, suggesting that it was formed by nonhomologous, i.e., random or illegitimate, recombination. In two of the recombinants, the 3' junction was apparently formed by homologous recombination, while in the other two recombinants, the 3' junction as well as the 5' junction might have involved a nonhomologous crossover. As reported previously, we found that the frequency of gene targeting increases monotonically with the length of the region of homology. Our results also indicate that targeting with fragments bearing one-sided homology can be as efficient as with fragments with bilateral homology, provided that the overall length of homology is comparable. The frequency of these events suggests that the immunoglobulin locus is particularly susceptible to nonhomologous recombination. Vectors designed for one-sided homologous recombination might be advantageous for some applications in genetic engineering.

Blotting, Southern↗

The relationship of angiotensin-converting enzyme gene to essential hypertension and drug treatment in Chongqing.

OBJECTIVE: To measure and discuss the association of the angiotensin converting enzyme (ACE) gene insertion/deletion(I/D) polymorphism with essential hypertension and drug treatment in Chongqing. METHODS: Polymerase chain reaction(PCR) method was used to amplify the sixteenth intron of the ACE gene for the analysis of 114 healthy controls and 75 patients with essential hypertension. At the same time, 49 patients with essential hypertension were randomly divided into two groups for a placebo-controlled crossover comparison of ACEI versus CCB. RESULTS: The frequency of homozygous alleles DD and the frequency of deletion alleles were significantly higher in the essential hypertension group than in the healthy controls(P>0.005). The blood pressure decreased more in the DD genotype subjects when treated with the ACEI but decreased less when treated with the CCB (15.91 vs 12.43, P<0.1). The blood pressure decreased more in the II genotype subjects when treated with the CCB but decreased less when treated with the ACEI (16.55 vs 9.6, P<0.05). CONCLUSION: The results indicate that a deletion (DD) polymorphism of the ACE gene is associated with essential hypertension in Chongqing. Detection of the ACE genotype in clinics is helpful to the selection of antihypertensive drugs.

Adult↗

Effect of cholestyramine on the symptoms of reflux gastritis. A randomized, double blind, crossover study.

Bile acids have been proposed to be important in the pathophysiology of the syndrome of "bile reflux gastritis" after surgery. To examine the role of cholestyramine, an ion exchange resin that binds bile acids, on symptoms of this syndrome, we did a randomized, double-blind crossover study on 16 patients. No differences in frequency of abdominal pain, nausea, vomiting, or bitter taste were observed among cholestyramine (4 g, three times daily for 3 weeks), placebo, and routine (dietary restriction and ad libitum antacid) treatment periods. We conclude that this regimen of cholestyramine was ineffective in symptomatic treatment of bile reflux gastritis.

Adult↗

The genetic analysis of recombination using adenovirus overlapping terminal DNA fragments.

We have studied the consequences of genetic recombination between overlapping terminal fragments of adenovirus genomes with respect to markers in the overlapping sequence. The findings are consistent with general recombination occurring approximately isotonically within the interval. In particular, single markers within the overlap, scored nonselectively, showed frequencies of recovery dependent on the position of their locus in relation to the ends of the overlap. Pairs of ts markers recombined to form ts+ progeny in proportion to their distance apart, provided the markers were oriented so that a single crossover between them would produce a full-length genome bearing both ts+ alleles. In the opposite orientation, where such a single crossover would be expected to produce ts/ts recombinants, the ts+ frequency was much lower, indicating that multiple recombination events are rare in this system. These findings rule out site-specific recombination, recombination occurring exclusively at the cleaved ends of the overlap, and recombination by means of mismatch repair of a heteroduplex the length of the overlap. They also indicate either that any heteroduplex junction region formed in the course of this reaction is quite short or that it is not subject to heteroduplex repair. Finally, our results demonstrate the efficacy of overlap recombination as a genetic and physical mapping tool and as a method of strain construction, and they suggest other applications, such as using overlap recombination to demonstrate that closely spaced pairs of markers (e.g., putative second-site reversions and their accompanying ts lesions) can be segregated.

Adenoviridae↗

Genetics of mouse antibodies. II. Recombination between VH genes and allotype.

The BAB/14 (BALB/c. C57BL/Ka-Ig-1b/HZ) congenic mouse strain was found to respond to (4-hydroxy-3-nitrophenyl)acetyl (NP)-chicken globulin immunization with nonheteroclitic anti-NP antibodies as does the BALB/c inbred partner strain. This is in contrast to the heteroclitic anti-NP response of the C57BL/Ka allotype donor strain and indicates that the allotype linked VH-NP gene was inherited with the VH-DEX gene from BALB/C rather than with the allotype genes from C57BL/Ka. This verifies the interpretation of BAB/14 as an immunoglobulin gene recombinant. Two recombinants between VH-DEX and Ig-1 allogype genes have been found among 554 chromosomes tested yielding a recombination frequency of 0.4%. Analysis of the BAB/14 recombinant showed that this crossover occurred between VH genes implying that VH genes and CH genes are not organized in separate clusters separated by a larger spacer region but are contiguous. Analysis of the recombination frequencies indicates that the total number of VH genes must be at least 50.

Animals↗

Microwave-induced thermal escape in Josephson junctions.

We investigate, by experiments and numerical simulations, thermal activation processes of Josephson tunnel junctions in the presence of microwave radiation. When the applied signal resonates with the Josephson plasma frequency oscillations, the switching current may become multivalued in a temperature range far exceeding the classical to quantum crossover temperature. Plots of the switching currents traced as a function of the applied signal frequency show very good agreement with the functional forms expected from Josephson plasma frequency dependencies on the bias current. Throughout, numerical simulations of the corresponding thermally driven classical Josephson junction model show very good agreement with the experimental data.

Journal Article↗

An Asp8Asn substitution results in the adenosine deaminase (ADA) genetic polymorphism (ADA 2 allozyme): occurrence on different chromosomal backgrounds and apparent intragenic crossover.

We have now determined the molecular genetic basis for the common biochemical polymorphism at the adenosine deaminase (ADA) locus. The ADA*2 allele contains a G to A transition at nt22 (relative to the ATG) that results in substitution of asparagine for aspartic acid at codon 8 (Asp8Asn). Introduction of the nucleotide substitution into an ADA 1 cDNA and transfection into monkey kidney (Cos) cells confirmed that the mutation resulted in expression of an enzyme that comigrated with the naturally occurring ADA 2 allozyme. The substitution of neutral asparagine for anionic aspartic acid is consistent with the more cathodal electrophoretic migration of ADA 2 as compared with ADA 1. The nucleotide substitution was found on at least two different genetic backgrounds, suggesting independent recurrence of the mutation. Consistent with independent recurrence, the G to A transition is at a CpG dinucleotide and represents a type of mutation that occurs with high frequency. We have also unexpectedly identified a probable intragenic crossover in the very large first intron that is rich in repetitive DNA sequences.

Adenosine Deaminase↗

Human otolith-ocular reflexes during off-vertical axis rotation: effect of frequency on tilt-translation ambiguity and motion sickness.

The purpose of this study was to examine how the modulation of tilt and translation otolith-ocular responses during constant velocity off-vertical axis rotation varies as a function of stimulus frequency. Eighteen human subjects were rotated in darkness about their longitudinal axis 30 degrees off-vertical at stimulus frequencies between 0.05 and 0.8 Hz. The modulation of torsion decreased while the modulation of horizontal slow phase velocity (SPV) increased with increasing frequency. It is inferred that the ambiguity of otolith afferent information is greatest in the frequency region where tilt (torsion) and translational (horizontal SPV) otolith-ocular responses crossover. It is postulated that the previously demonstrated peak in motion sickness susceptibility during linear accelerations around 0.3 Hz is the result of frequency segregation of ambiguous otolith information being inadequate to distinguish between tilt and translation.

Adult↗

Reciprocal and nonreciprocal recombination in diploid clones from Bacillus subtilis protoplast fusion: Association with the replication origin and terminus.

The primary heterodiploid bacteria regenerated after Bacillus subtilis fusion, although generally noncomplementing diploids, behave in pedigree analysis as multipotential systems. Individual diploid colonies yielding complete reciprocal recombinant (RR) progeny-often accompanied by one or both parents-constitute 10-30% of the total recombinant-forming units. The RR (reciprocal for 8-11 genes) usually occur in equivalent numbers both among and within individual colonies. Novel for bacteria, they demonstrate that entire parental genomes brought together within a diploid protoplast are retained as two independent replicons able to undergo classical recombination characteristic of eukaryotic gametogenesis. Parental or recombinant genomes are also subject to multiple rounds of recombination without obligate segregation and often not reciprocal. Diploid recombinant clones, sharing streptomycin resistance but reciprocal for auxotrophic markers, have displayed a partial ability to make a facultative shift in chromosome expression. They have also produced two types of prototrophs: a stable one (presumably haploid and recombinant) and an unstable one, (diploid and temporarily complementing at low frequency). It follows that chromosome extinction may affect both parental and recombinant chromosomes and does not interfere with recombination. Analysis of the number and chromosomal distribution of crossovers in all recombinants and those from single diploid clones shows increased frequency of exchange in the regions of the replication origin and terminus, possibly a result of the association of these sites with the cell wall or membrane.

Journal Article↗

A chromosome marker for the early detection of mouse embryos carrying the neural tube defect mutation splotch.

A major problem in the study of neural tube defects caused by the splotch (Sp) gene in the mouse has been the identification of gene carriers or potentially affected embryos at an early stage of development, since the gene's effects become visible only late in gestation or after birth. To aid in the identification of Sp carriers, we have developed a technique using a Robertsonian translocation as a marker for this gene. The accuracy of identification is reduced by crossing-over between the Sp locus and the centromere but, because of crossover suppression in the particular cross used, there was only 23.2% recombination compared with the known map distance of 36%. Paternal age had no effect on the frequency of recombination, but individual males differed significantly in the degree of crossover suppression.

Animals↗

Crossover interference in Saccharomyces cerevisiae requires a TID1/RDH54- and DMC1-dependent pathway.

Two RecA-like recombinases, Rad51 and Dmc1, function together during double-strand break (DSB)-mediated meiotic recombination to promote homologous strand invasion in the budding yeast Saccharomyces cerevisiae. Two partially redundant proteins, Rad54 and Tid1/Rdh54, act as recombinase accessory factors. Here, tetrad analysis shows that mutants lacking Tid1 form four-viable-spore tetrads with levels of interhomolog crossover (CO) and noncrossover recombination similar to, or slightly greater than, those in wild type. Importantly, tid1 mutants show a marked defect in crossover interference, a mechanism that distributes crossover events nonrandomly along chromosomes during meiosis. Previous work showed that dmc1Delta mutants are strongly defective in strand invasion and meiotic progression and that these defects can be partially suppressed by increasing the copy number of RAD54. Tetrad analysis is used to show that meiotic recombination in RAD54-suppressed dmc1Delta cells is similar to that in tid1; the frequency of COs and gene conversions is near normal, but crossover interference is defective. These results support the proposal that crossover interference acts at the strand invasion stage of recombination.

Cell Cycle Proteins↗

Recql5 and Blm RecQ DNA helicases have nonredundant roles in suppressing crossovers.

In eukaryotes, crossovers in mitotic cells can have deleterious consequences and therefore must be suppressed. Mutations in BLM give rise to Bloom syndrome, a disease that is characterized by an elevated rate of crossovers and increased cancer susceptibility. However, simple eukaryotes such as Saccharomyces cerevisiae have multiple pathways for suppressing crossovers, suggesting that mammals also have multiple pathways for controlling crossovers in their mitotic cells. We show here that in mouse embryonic stem (ES) cells, mutations in either the Bloom syndrome homologue (Blm) or the Recql5 genes result in a significant increase in the frequency of sister chromatid exchange (SCE), whereas deleting both Blm and Recql5 lead to an even higher frequency of SCE. These data indicate that Blm and Recql5 have nonredundant roles in suppressing crossovers in mouse ES cells. Furthermore, we show that mouse embryonic fibroblasts derived from Recql5 knockout mice also exhibit a significantly increased frequency of SCE compared with the corresponding wild-type control. Thus, this study identifies a previously unknown Recql5-dependent, Blm-independent pathway for suppressing crossovers during mitosis in mice.

Adenosine Triphosphatases↗

Comparative studies of gastrointestinal tolerance and acceptability of milk chocolate containing either sucrose, isomalt or sorbitol in healthy consumers and type II diabetics.

The objective was to compare reaction of adult consumers of confectionery to milk chocolate made with either isomalt, sucrose or sorbitol. Test chocolate was eaten by subjects at home during 7 days in amounts chosen by them up to a maximum of 100 g per day. In a double-blind crossover trial isomalt chocolate was associated in healthy consumers (n = 58) with increased motion frequency, wind and flatulence compared with sucrose chocolate. However, the intensity of these gastrointestinal effects was predominantly slight and insufficient to affect acceptability. In separate crossover trials, reactions of Type II diabetic consumers to eating isomalt chocolate (n = 53) or sorbitol chocolate (n = 51) were compared to reactions when eating no chocolate. Both isomalt and sorbitol chocolate were associated with higher incidence of wind and flatulence than for no chocolate, but only sorbitol chocolate increased motion frequency. Again intensity of gastrointestinal effects was slight. It is concluded that isomalt has potential use in both regular and diabetic chocolate.

Abdominal Pain↗

The effect of two toothpaste detergents on the frequency of recurrent aphthous ulcers.

The aim of the present clinical double-blind crossover study was to investigate the effect of two different toothpaste detergents, sodium lauryl sulfate (SLS) and cocoamidopropyl betaine (CAPB), as compared with a detergent-free paste, on 30 patients with frequent occurrences of recurrent aphthous ulcers. The study consisted of three 6-week periods during which the patients brushed twice daily with the different test toothpastes. The localization and number of new ulcers were assessed. A significantly higher frequency of aphthous ulcers was demonstrated when the patients brushed with an SLS- than with a CAPB-containing or a detergent-free placebo paste. An SLS-free toothpaste may thus be recommended for patients with recurrent aphthous ulcers.

Adolescent↗

Tam1, a telomere-associated meiotic protein, functions in chromosome synapsis and crossover interference.

The TAM1 gene of Saccharomyces cerevisiae is expressed specifically during meiosis and encodes a protein that localizes to the ends of meiotic chromosomes. In a tam1 null mutant, there is an increase in the frequency of chromosomes that fail to recombine and an associated increase in homolog nondisjunction at meiosis I. The tam1 mutant also displays an increased frequency of precocious separation of sister chromatids and a reduced efficiency of distributive disjunction. The defect in distributive disjunction may be attributable to overloading of the distributive system by the increased number of nonrecombinant chromosomes. Recombination is not impaired in the tam1 mutant, but crossover interference is reduced substantially. In addition, chromosome synapsis is delayed in tam1 strains. The combination of a defect in synapsis and a reduction in interference is consistent with previous studies suggesting a role for the synaptonemal complex in regulating crossover distribution. tam1 is the only known yeast mutant in which the control of crossover distribution is impaired, but the frequency of crossing over is unaffected. We discuss here possibilities for how a telomere-associated protein might function in chromosome synapsis and crossover interference.

Chromosome Mapping↗

Critical light scattering in liquids

We compare theoretical results for the characteristic frequency of the Rayleigh peak calculated in one-loop order within the field theoretical method of the renormalization group theory with experiments and other theoretical results. Our expressions describe the nonasymptotic crossover in temperature, density, and wave vector. In addition we discuss the frequency dependent shear viscosity evaluated within the same model and compare our theoretical results with recent experiments in microgravity.

Journal Article↗