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PubMed2002

An initiation site for meiotic crossing-over and gene conversion in the mouse.

During meiosis, the reductional segregation of homologous chromosomes at the first meiotic division requires reciprocal exchange (crossing over) between homologs. The number of crossovers is tightly regulated (one to two per homolog in mice), and their distribution in the genome is not random-recombination 'hot' and 'cold' regions can be identified. We developed a molecular assay to study these events directly in mouse germ cells. This analysis was developed with reference to the proteosome subunit beta type 9 (Psmb9, previously called Lmp2) hot-spot region identified through genetic analysis. Here we show that this hot spot is an initiation site of meiotic recombination on the basis of two observations: (i) crossover density is maximal in an interval of 210 bp and decreases on both sides of this region; (ii) a high frequency of gene conversion is found in the region of highest crossover density. We then used this strategy to carry out the first temporal analysis of meiotic recombination in mouse spermatogenesis and demonstrate that crossover events occur during the pachytene stage of meiotic prophase.

Animals↗
PubMed1985

Methodological aspects of drug trials in migraine.

Treatment of migraine attacks and prophylactic treatment of migraine are each discussed under the four headings: patient selection, trial design, evaluation of results and statistics. Checklist of problems encountered in these trials are given in the tables. The unsuitability of currently used migraine definitions for scientific investigations is stressed. Operational diagnostic criteria for common and classic migraine are given. No clear separation of common migraine attacks and interval headaches is possible, but the problem can be reduced by setting an upper limit (4-6/month) on migraine attack frequency. For the treatment of migraine attacks, the crossover design should always be used. We suggest dose-response studies to solve the problem of equipotency of doses, when 2 drugs are compared. A prophylactic drug should be studied both with the crossover design and with the less powerful group comparison design. Evaluation of results should be based on patients' attack report forms and, in prophylactic studies, a headache diary. We suggest global rating of attack severity. A rather simple headache index (sum of severity scores for each day with migraine) and perhaps a sum of global scores should be used in prophylactic trials. Confidence limits and power should be given, particularly when statistically insignificant results are reported. Migraine frequency often decreases with time regardless of treatment, and this 'time effect' should be separated from the therapeutic effect by appropriate statistical methods.

Analgesics↗
PubMed1989

Effects of changes in frequency of reticular contractions on fluid and particulate passage rates in cattle.

Four ruminally cannulated Hereford steers (531 +/- 32 kg) without weights (control; C) or with 24-kg weights (W) in the rumen were used in a double crossover design experiment to determine the effects of changes in frequency of reticular contractions at maintenance feed intakes on passage rates of particulate matter and fluid from the rumen. Duration of reticular contractions, apparent digestibilities and heat and methane productions were also determined. Weights in the rumen had no effect (P = .07) on frequency of reticular contractions before feeding; however, frequencies of reticular contractions were higher in C than in W steers during feeding (P = .03) and after feeding (P = .05). Weights in the rumen increased the average duration of contractions before feeding (P = .04), during feeding (P = .01) and after feeding (P = .05) by 12, 15 and 15%, respectively. Fractional outflow rates of ruminal and whole gastrointestinal particulate matter were 38% and 49% lower (P = .001) in C than in W steers, respectively. Weights in the rumen reduced (P = .001) methane production (liters/d) by 29%. Methane production was inversely correlated with fractional outflow rate for ruminal particulate matter (r = -.53; P = .034). Heat productions for W and C steers were 12.3 and 12.6 Mcal/d, respectively. Weights in the rumen had no effect on apparent digestibility. Duration of reticular contractions, in contrast to frequency of contractions, was the important factor influencing passage rates of both ruminal fluid and particulate matter. Methane production was decreased when passage rates were increased.(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed↗
PubMed1985

Meiotic recombination between duplicated genetic elements in Saccharomyces cerevisiae.

We have studied the meiotic recombination behavior of strains carrying two types of duplications of an 18.6-kilobase HIS4 Bam HI fragment. The first type is a direct duplication of the HIS4 Bam HI fragment in which the repeated sequences are separated by Escherichia coli plasmid sequences. The second type, a tandem duplication, has no sequences intervening between the repeated yeast DNA. The HIS4 genes in each region were marked genetically so that recombination events between the duplicated segments could be identified. Meiotic progeny of the strains carrying the duplication were analyzed genetically and biochemically to determine the types of recombination events that had occurred. Analysis of the direct vs. tandem duplication suggests that the E. coli plasmid sequences are recombinogenic in yeast when homozygous. In both types of duplications recombination between the duplicated HIS4 regions occurs at high frequency and involves predominantly interchromosomal reciprocal exchanges (equal and unequal crossovers). The striking observation is that intrachromosomal reciprocal recombination is very rare in comparison with interchromosomal reciprocal recombination. However, intrachromosomal gene conversion occurs at about the same frequency as interchromosomal gene conversion. Reciprocal recombination events between regions on the same chromatid are the most infrequent exchanges. These data suggest that intrachromosomal reciprocal exchanges are suppressed.

Chromosomes↗
PubMed2023

Comparative transmission genetics of introgressed chromatin in reciprocal advanced backcross populations in Gossypium (cotton) polyploids.

Introgression is a potential source of valuable genetic variation and interspecific introgression lines are important resources for plant breeders to access novel alleles. Experimental advanced-generation backcross populations contain individuals with genomic compositions similar to those resulting from natural interspecific hybridization and provide opportunities to study the nature and transmission pattern of donor chromatin in recipient genomes. Here, we analyze transmission of donor chromatin in reciprocal backcrosses between G. hirsutum and G. barbadense. Across the genome, recurrent backcrossing in both backgrounds yielded donor chromatin at slightly higher frequencies than the Mendelian expectation in BC5F1 plants, while the average frequency of donor alleles in BC5F2 segregating families was less than expected. In the two subgenomes of polyploid cotton, the rate of donor chromatin introgression was similar. Although donor chromatin was tolerated over much of the recipient genomes, 21 regions recalcitrant to donor alleles were identified. Only limited correspondence is observed between the recalcitrant regions in the two backgrounds, suggesting the effect of species background on introgression of donor segments. Genetic breakdown was progressive, with floral abscission and seed inviability ongoing during backcrossing cycles. Regions of either high or low introgression tended to be in terminal chromosomal regions that are generally rich in both genes and crossover events, with long stretches around the centromere having limited crossover activity resulting in relatively constant low introgression frequencies. Constraints on fixation and selection of donor alleles highlights the challenges of utilizing introgression breeding in crop improvement.

Humans↗
PubMed2000

Rheological properties of ice cream mixes and frozen ice creams containing fat and fat replacers.

Ice cream mixes and frozen ice creams at milk fat levels of 12%, 8%, 6%, 6% plus a protein-based fat replacer, and 6% plus a carbohydrate-based fat replacer were evaluated for viscoelastic properties by dynamic testing with sinusoidal oscillatory tests at various frequencies. The storage modulus (G'), loss modulus (G"), and tan delta (G"/G') were calculated for all the treatments to determine changes in the viscous and elastic properties of the mixes and frozen ice creams due to fat content. In ice cream mixes, G' and G" exhibited a strong frequency dependence. The G" was higher than G' throughout the frequency range (1 to 8 Hz) examined, without any crossover, except for the 12% mix. Elastic properties of the ice cream mixes decreased as fat content decreased. Tan delta values indicated that fat replacers did not enhance the elastic properties of the ice cream mixes. In all frozen ice creams, G' and G" again showed a frequency dependence throughout the range tested (0.5 to 10 Hz). The amount of fat in ice creams and the degree of fat destabilization affected the elasticity in the frozen product. Even though the ice creams did not have significant elastic properties, when compared as a group the samples with higher fat content had higher elastic properties. The addition of protein-based and carbohydrate-based fat replacers did not enhance the elastic properties of the ice creams but did increase the viscous properties.

Chemical Phenomena↗
PubMed2005

Homologous crossovers among molecules of brome mosaic bromovirus RNA1 or RNA2 segments in vivo.

Previously we demonstrated frequent homologous crossovers among molecules of the RNA3 segment in the tripartite brome mosaic bromovirus (BMV) RNA genome (A. Bruyere, M. Wantroba, S. Flasinski, A. Dzianott, and J. J. Bujarski, J. Virol. 74:4214-4219, 2000). To further our knowledge about mechanisms of viral RNA genome variability, in this paper we have studied homologous recombination in BMV RNA1 and RNA2 components during infection. We have found that basal RNA-RNA crossovers could occur within coding regions of both RNAs, although recombination frequencies slightly varied at different RNA sections. In all cases, the frequencies were much lower than the rate observed for the intercistronic recombination hot spot in BMV RNA3. Probability calculations accounted for at least one homologous crossover per RNA molecule per replication cycle. In addition, we have demonstrated an efficient repair of mutations within the conserved 3' and 5' noncoding regions, most likely due to error-prone BMV RNA replication. Overall, our data verify that homologous crossovers are common events a during virus life cycle, and we discuss their importance for viral RNA genetics.

Bromovirus↗
PubMed2001

Calcium-channel blockers for Raynaud's phenomenon in systemic sclerosis.

OBJECTIVE: Most patients with systemic sclerosis (SSc) have Raynaud's phenomenon (RP), which is often more severe than idiopathic RP. This study was a meta-analysis to determine the efficacy of calcium-channel blockers for the treatment of RP in SSc. The primary outcome measures were frequency and severity of ischemic attacks, digital skin temperature, patient and physician global assessments, and digital ulcers. METHODS: The Cochrane search strategy was used to ascertain all trials in all languages. Primary data sources included Medline, Current Contents, and the Cochrane Controlled Trials Register. Studies that met the inclusion criteria were randomized controlled trials of >2 days' duration with a dropout rate of <35%. Twenty-nine studies were found, of which 8 randomized controlled trials were eligible for inclusion. The total number of patients included was small (n = 109). Most trials included primary and secondary RP, and the main reasons for trial exclusion were inability to extract subset data on SSc patients (18 trials), data published previously (2 trials), and lack of a control group (1 trial). Data were abstracted independently by 2 reviewers, and either a weighted mean difference (WMD) or a standardized mean difference (SMD) was calculated for all continuous outcomes; however, information was not available for all outcomes within trials. RESULTS: The WMD of all calcium-channel blockers versus placebo (6 trials) and of nifedipine alone versus placebo (5 trials) for the reduction in the frequency of ischemic attacks over a 2-week period was -8.31 (95% confidence interval [95% CI] -15.71, -0.91) and -10.21 (95% CI -20.09, -0.34), respectively. The SMD of all calcium-channel blockers versus placebo (3 trials) and of nifedipine alone versus placebo (2 trials) for the reduction in the severity of ischemic attacks was -0.69 (95% CI -1.21, -0.17) and -0.99 (95% CI -1.74, -0.24), respectively. CONCLUSION: Calcium-channel blockers for RP in SSc have been tested in several small clinical trials and appear to lead to significant clinical improvement in both the frequency and the severity of ischemic attacks. Most trials were crossover trials in which order effect was not studied. This could have introduced bias. The results of this study suggest that the efficacy of calcium-channel blockers in reducing the severity and frequency of ischemic attacks in RP secondary to SSc is moderate at best (mean reduction of 8.3 attacks in 2 weeks and 35% less severity), and a further large, randomized controlled trial needs to be conducted.

Calcium Channel Blockers↗
PubMed1997

Controlled trials of inositol in psychiatry.

Inositol is a simple polyol precursor in a second messenger system important in the brain. Cerebrospinal fluid inositol has been reported as decreased in depression. A double-blind controlled trial of 12 g daily of inositol in 28 depressed patients for four weeks was performed. Significant overall benefit for inositol compared to placebo was found at week 4 on the Hamilton Depression Scale. No changes were noted in hematology, kidney or liver function. Since many antidepressants are effective in panic disorder, twenty-one patients with panic disorder with or without agoraphobia completed a double-blind, placebo-controlled, four week, random-assignment crossover treatment trial of inositol 12 g per day. Frequency and severity of panic attacks and severity of agoraphobia declined significantly with inositol compared to placebo. Side-effects were minimal. Since serotonin re-uptake inhibitors benefit obsessive compulsive disorder (OCD) and inositol is reported to reverse desensitization of serotonin receptors, thirteen patients with OCD completed a double-blind controlled crossover trial of 18 g inositol or placebo for six weeks each. Inositol significantly reduced scores of OCD symptoms compared with placebo. A controlled double-blind crossover trial of 12 g daily of inositol for a month in twelve anergic schizophrenic patients, did not show any beneficial effects. A double-blind controlled crossover trial of 6 g of inositol daily vs. glucose for one month each was carried out in eleven Alzheimer patients, with on clearly significant therapeutic effects. Antidepressant drugs have been reported to improve attention deficit disorder (ADDH) with hyperactivity symptomatology. We studied oral inositol in children with ADDH in a double-blind, crossover, placebo-controlled manner. Eleven children, mean age 8.9 +/- 3.6 years were enrolled in an eight week trial of inositol or placebo at a dose of 200 mg/kg body weight. Results show a trend for aggravation of the syndrome with myo-inositol as compared to placebo. Recent studies suggest that serotonin re-uptake inhibitors are helpful in at least some symptoms of autism. However a controlled double-blind crossover trial of inositol 200 mg/kg per day showed no benefit in nine children with autism. Cholinergic agonists have been reported to ameliorate electroconvulsive therapy (ECT)-induced memory impairment. Inositol metabolism is involved in the second messenger system for several muscarinic cholinergic receptors. Inositol 6 g daily was given in a crossover-double-blind manner for five days before the fifth or sixth ECT to a series of twelve patients, without effect. These results suggest that inositol has therapeutic effects in the spectrum of illness responsive to serotonin selective re-uptake inhibitors, including depression, panic and OCD, and is not beneficial in schizophrenia, Alzheimer's ADDH, autism or ECT-induced cognitive impairment.

Adult↗
PubMed1999

Crossover accelerates evolution in GAs with a Babel-like fitness landscape: mathematical analyses.

The effectiveness of crossover in accelerating evolution in genetic algorithms (GAs) is studied with a haploid finite population of bit sequences. A Babel-like fitness landscape is assumed. There is a single bit sequence (schema) that is significantly more advantageous than all the others. We study the time until domination of the advantageous schema (&Tau;&subd;). Evolution proceeds with appearance, spread, and domination of the advantageous schema. The most important process determining &Tau;&subd; is the appearance (creation) of the advantageous schema. Crossover helps this creation process and enhances the rate of evolution. To study this effect, we first establish an analytical method to estimate &Tau;&subd; with or without crossover. Then, we conduct a numerical analysis using the frequency vector representation of the population with the recurrence relations formulated after GA operations. Finally, we carry out direct computer simulations with simple GAs operating on a population of binary strings directly prepared in the computer memory to examine the performance of the two analytical methods. It is shown that &Tau;&subd; is reduced greatly by crossover with a mildly high rate when the mutation rate is adjusted to a moderate value and that an advantageous schema has a fairly larger order (the number of bits). From these observations, we can determine implementation criteria for GAs, which are useful when we are applying GAs to engineering problems having a conspicuously discontinuous fitness landscape.

Algorithms↗
PubMed2002

Alternate site glucose testing: a crossover design.

A crossover design study was conducted to determine if reducing pain, by using alternative sites off the finger tip, would increase testing frequency and improve clinical outcome as measured by hemoglobin A1c (HbA1c). Subjects with type 1 and type 2 diabetes tested with the FreeStyle meter (group 1) or tested with their original meter (group 2). After three months the subjects used the alternate meter. Testing frequency and blood glucose concentrations were recorded for the month before the study began and monthly thereafter. HbA1c was tested initially, at the crossover point, and at study conclusion. Insulin users increased testing frequency from 2.4 to 3.0 tests/day. Testing frequency for non-insulin users remained the same at 1.5 tests/day. Testing frequency was the same with the FreeStyle and the original meters. The average HbA1c was 7.4% (SD 1.5%) initially, 7.3% (SD 1.5%) at the crossover point, and 6.9% (SD 1.1%) at study conclusion. There was no significant difference in HbA1c measurements between meter types. Preference rankings were 76% for FreeStyle, 20% for their original meter, and 4% preferred both meters equally. This population tended to be in good glycemic control with 70% having HbA1c at < or = 8.0 at study initiation. Subjects preferred testing with the FreeStyle meter (76%), but did not increase testing frequency. Study participants tended to be in good testing compliance and glycemic control with little room for improvement. A population in poor glucose control might show differences in outcome with off site monitoring.

Adult↗
PubMed2004

Effects of a single bout of exercise on resting heart rate variability.

PURPOSE: Chronic exercise training has been shown to have a positive influence on cardiac autonomic function as assessed by measures of heart rate variability (HRV). Recent evidence indicates that several benefits associated with exercise training (e.g., improved insulin action, reduced blood pressure, improved blood lipid profile) may be realized transiently after a single bout of exercise. As many of these effects of recent exercise are linked to cardiovascular control systems, the purpose of this investigation was to test the hypothesis that a single bout of exercise would result in favorable changes in cardiac autonomic function as assessed by frequency-domain measures of HRV. METHODS: Subjects were 11 healthy male volunteers ages 18-35 yr. Resting HRV measures were obtained during 5 min of paced breathing before and 1, 3, 6, and 22 h after a 60-min bout of cycling exercise at approximately 65% of peak oxygen uptake. Identical measures were obtained in a nonexercise condition based on a randomized crossover design. RESULTS: Exercise resulted in increased high-frequency HRV, decreased low-frequency HRV, and consequently a decrease in the ratio of low-frequency to high-frequency HRV compared with the nonexercise condition. Additionally, a time-domain measure of HRV (pNN50) was markedly higher in the exercise condition as compared to nonexercise. CONCLUSION: The changes in cardiac autonomic function observed are similar to those seen in investigations of long-term training. These changes indicate a shift in autonomic function toward increased parasympathetic nervous system activity and decreased sympathetic nervous system activity, suggesting a more stable autonomic environment for the heart. These results may provide further evidence of the cardioprotective effects of a single bout of submaximal exercise.

Adolescent↗
PubMed2005

REC, Drosophila MCM8, drives formation of meiotic crossovers.

Crossovers ensure the accurate segregation of homologous chromosomes from one another during meiosis. Here, we describe the identity and function of the Drosophila melanogaster gene recombination defective (rec), which is required for most meiotic crossing over. We show that rec encodes a member of the mini-chromosome maintenance (MCM) protein family. Six MCM proteins (MCM2-7) are essential for DNA replication and are found in all eukaryotes. REC is the Drosophila ortholog of the recently identified seventh member of this family, MCM8. Our phylogenetic analysis reveals the existence of yet another family member, MCM9, and shows that MCM8 and MCM9 arose early in eukaryotic evolution, though one or both have been lost in multiple eukaryotic lineages. Drosophila has lost MCM9 but retained MCM8, represented by REC. We used genetic and molecular methods to study the function of REC in meiotic recombination. Epistasis experiments suggest that REC acts after the Rad51 ortholog SPN-A but before the endonuclease MEI-9. Although crossovers are reduced by 95% in rec mutants, the frequency of noncrossover gene conversion is significantly increased. Interestingly, gene conversion tracts in rec mutants are about half the length of tracts in wild-type flies. To account for these phenotypes, we propose that REC facilitates repair synthesis during meiotic recombination. In the absence of REC, synthesis does not proceed far enough to allow formation of an intermediate that can give rise to crossovers, and recombination proceeds via synthesis-dependent strand annealing to generate only noncrossover products.

Animals↗
PubMed1987

Effect of diltiazem and propranolol on myocardial ischaemia during unrestricted daily life in patients with effort-induced chronic stable angina pectoris.

To examine the effect of diltiazem 60 mg thrice daily and propranolol 80 mg thrice daily on myocardial ischaemia during unrestricted daily life, we have studied 14 patients with established effort-induced chronic stable angina pectoris in a double-blind crossover study. Ambulatory electrocardiographic monitoring was performed using frequency modulated (FM) tape recorder after 2 weeks of placebo therapy and at the end of each 4 week treatment period for a minimum of 24 h. The mean (+/- SEM) number of episodes of ST-segment depression greater than 1 mm during placebo treatment were 97 +/- 28, and these fell to 54 +/- 27 during diltiazem treatment (p less than 0.05) and to 12 +/- 6 during propranolol treatment (p less than 0.02). The maximal depth of ST-segment depression in 24 h, which indicates the severity of the episode, was 3.3 +/- 0.4 mm during placebo, 2.5 +/- 0.2 mm during diltiazem (p less than 0.05), and 1.6 +/- 0.5 mm during propranolol (p less than 0.01). Both diltiazem and propranolol treatment produced significant reduction in the total area of ST-segment depression observed during 24 h. A uniform reduction in the mean heart rate during the 24 h was observed during propranolol therapy and not during diltiazem therapy. Both diltiazem and propranolol treatment improved indices of myocardial ischaemia during daily normal unrestricted life, as measured by ambulatory ST-segment monitoring in patients with established chronic stable angina pectoris. Their differing effects on heart rate suggest that they act by different mechanisms.

Aged↗
PubMedDate not supplied

Acute vagus nerve stimulation does not suppress spike and wave discharges in genetic absence epilepsy rats from Strasbourg.

We evaluated the efficacy of vagus nerve stimulation (VNS) in Genetic Absence Epilepsy Rats from Strasbourg (GAERS), a validated model for absence epilepsy. In the first experiment, we investigated whether VNS applied at seizure onset can interrupt spike and wave discharges (SWD). In the second experiment, we investigated whether SWD are suppressed or shortened in duration when VNS is applied several hours per day. Both control and VNS groups underwent EEG and VNS electrode implantation. For the first experiment, a randomized crossover design was used. Stimuli (amplitude: 3 V; frequency: 30 Hz; pulse duration: 500 micros) were given when an SWD occurred on the EEG. The experiment was repeated the next day. In the second experiment, treated animals were stimulated (amplitude: 1.5 mA; frequency: 30 Hz; pulse duration: 500 micros; on/off time cycle: 30 s / 5 min) for 3h per day, during five consecutive days. In the first experiment, the duration of the SWD was increased on day 1, (P < 0.05). There was no difference in SWD duration on Day 2. In the second experiment, no significant differences could be found in number, duration and EEG frequency of SWD. VNS applied at the onset of an SWD can prolong the duration of SWD in GAERS. As a 5-day stimulation protocol had no effect, long-term VNS might be necessary to affect SWD.

Action Potentials↗
PubMed2000

Frequent ectopic recombination of virulence factor genes in telomeric chromosome clusters of P. falciparum.

Persistent and recurrent infections by Plasmodium falciparum malaria parasites result from the ability of the parasite to undergo antigenic variation and evade host immune attack. P. falciparum parasites generate high levels of variability in gene families that comprise virulence determinants of cytoadherence and antigenic variation, such as the var genes. These genes encode the major variable parasite protein (PfEMP-1), and are expressed in a mutually exclusive manner at the surface of the erythrocyte infected by P. falciparum. Here we identify a mechanism by which var gene sequences undergo recombination at frequencies much higher than those expected from homologous crossover events alone. These recombination events occur between subtelomeric regions of heterologous chromosomes, which associate in clusters near the nuclear periphery in asexual blood-stage parasites or in bouquet-like configurations near one pole of the elongated nuclei in sexual parasite forms. We propose that the alignment of var genes in heterologous chromosomes facilitates gene conversion and promotes the diversity of antigenic and adhesive phenotypes. The association of virulence factors with a specific nuclear subcompartment may also have implications for variation during mitotic recombination in asexual blood stages.

Animals↗
PubMed1980

Triplicated alpha-globin loci in humans.

We have identified 12 individuals who are heterozygous for a chromosome with three alpha-globin genes. We determined the presence of the third alpha-globin locus by restriction endonuclease digestion and hybridization with alpha-globin cDNA probes. The three alpha-globin loci resided in an elongated fragment on digestion with EcoRI, BamHI, and Xba I, and the third locus was present in an additional 3.7-kilobase fragment on digestion with Hpa I, Sac I, and Bgl II. The locations of the restriction sites are compatible with a nonhomologous crossover mechanism producing the triple alpha loci. The frequency of the triple alpha loci was 0.0036 in American blacks and less than 0.004 in Sardinians, but was higher in Greek Crypriots (0.05). Individuals with five alpha-globin genes (alpha alpha/alpha alpha alpha) apparently showed no clinical or hematologic abnormalities.

Black People↗
PubMed2004

Isotropic convection scenarios in an anisotropic fluid.

We study a new variant of electroconvection using a homeotropically aligned nematic liquid crystal. The novelty of this system is a direct transition to roll- or square-type patterns controlled by the frequency of the applied voltage with a rich crossover scenario and strong influence of the zigzag instability even at onset. From the weakly nonlinear theory and simulations of an adapted Swift-Hohenberg model one can understand essential features of the phase diagram. In particular we find a quasiperiodic pattern with square symmetry.

Journal Article↗