Rhythm-Specific Mutations in Drosophila rajasekari Altered Adult Locomotor Activity Rhythm but Not Eclosion Rhythm

The early and late strains for phase angle difference (Φ) of adult locomotor activity in Drosophila rajasekari were developed by artificial selection; these strains differed in Φ, activity pattern, activity level, free-running period (τ) in constant darkness (DD) and light induced phase shifts from those of the wild type (Joshi, 1998). The present studies were designed to determine whether or not the psi-mutations for adult locomotor activity rhythm had also altered the fundamental properties of the eclosion rhythms in these strains. The circadian rhythms of eclosion have been studied in the wild type, the early and late strains. In contrast to the effects on the locomotor activity rhythms in the early and late strains, the psi-mutations have no apparent effect on the eclosion median in light-dark cycles of 12 : 12 h, on τ in DD, light induced phase shifts or subjective light sensitivity in these strains. Thus the psi-mutations for the adult locomotor activity rhythms in D. rajasekari appear to be rhythm-specific mutations altering the locomotor rhythms but not the eclosion rhythms.     

Investigation of Natural Genetic Variation in the Circadian System of Drosophila melanogaster. II. Biometrical Analyses of Locomotor Activity Rhythms Recorded in Constant Darkness

The locomotor activity rhythms of 24 isochromosomal strains of Drosophila melanogaster were recorded in constant conditions, using an experimental design suitable for the serial screening with so many strains using limited equipment. The data obtained were subjected to biometrical genetic analysis. The results show that four characteristics of the rhythms (period, phase, definition, and waveform) display genetically based variation in expression and that each appears to be affected by a range of genes.     

Complementarity of thermoregulatory rhythms in Micropterus salmoides and M. dolomieui

Largemouth (Micropterus salmoide) and smallmouth (M. dolomieui) blackbasses tested in an electronic shuttlebox exhibited behavioral thermoregulatory rhythms which were temporally complementary. With a LD 12 : 12 photoperiod, M. dolomieui exhibited a preferred-temperature peak of 30.1°C during the latter portion of the photophase, when M. salmoides reached a minimum of 27.1°C. M. dolomieui exhibited a minimum of 26.6°C during the latter portion of scotophase, while M. salmoides remained at a significantly higher plateau of about 29°C, with a peak of 29.5°C at the midpoint of scotophase. The phase relations of the thermoregulatory rhythms relative to photoperiod suggest that they are endogenously timed circadian rhythms entrained by photoperiod. The thermotemporal complementarity of these rhythms suggests an aspect of niche segregation between these largely sympatric congeneric species.     

A Circadian Rhythm in Mice that is Unaffected by the Period Mutation at Clock

Circadian rhythms of locomotor behavior were induced in mice using chronic d-amphetamine. The periods of the rhythms at various doses of amphetamine were unaffected by a mutation at the clock locus that lengthens circadian period. Amphetamine-induced rhythms were superimposed on the hostÕs natural circadian rhythm that is driven by the pacemaker in the suprachiasmatic nucleus. The results demonstrate the existence of an alternative mechanism for generating circadian rhythms that does not require the activity of canonical clock genes.     

Rhythm-Specific Mutations in Drosophila rajasekari Altered Adult Locomotor Activity Rhythm but Not Eclosion Rhythm

The early and late strains for phase angle difference (Φ) of adult locomotor activity in Drosophila rajasekari were developed by artificial selection; these strains differed in Φ, activity pattern, activity level, free-running period (τ) in constant darkness (DD) and light induced phase shifts from those of the wild type (Joshi, 1998). The present studies were designed to determine whether or not the psi-mutations for adult locomotor activity rhythm had also altered the fundamental properties of the eclosion rhythms in these strains. The circadian rhythms of eclosion have been studied in the wild type, the early and late strains. In contrast to the effects on the locomotor activity rhythms in the early and late strains, the psi-mutations have no apparent effect on the eclosion median in light-dark cycles of 12 : 12 h, on τ in DD, light induced phase shifts or subjective light sensitivity in these strains. Thus the psi-mutations for the adult locomotor activity rhythms in D. rajasekari appear to be rhythm-specific mutations altering the locomotor rhythms but not the eclosion rhythms.     

Circadian Locomotor Activity in the Larva and Adult Fall Armyworm,Spodoptera frugiperda (Noctuidae): Effect of Feeding with the Resistant Variety of Maize CML67

The circadian rhythms of locomotor activity were studied in larvae and adult Fall Armyworms, Spodoptera frugiperda, fed a nutritional diet (control) or a diet containing the maize variety CML67 (experimental). Activity was monitored using infrared-light crossings and rhythms were evaluated using actograms, average waveform, and X² periodograms. Results show that larvae grown on the experimental diet did not display conspicuous circadian rhythms in constant darkness before pupation and showed poor responses to light-dark cycles. Significant differences between the two groups were observed in their maximal activity and activity-rest ratios. Adults fed either diet displayed circadian rhythms of locomotor activity, however, differences were still found in their activity-rest ratios. Results obtained indicate that animals fed with diet containing the maize variety CLM67, have significant differences in the expression of circadian locomotor activity in larvae under constant darkness and in their responses to artificial light-dark conditions, suggesting that the maize variety CML67 may possess some active substance(s) that affect the maturation of the circadian system, controlling locomotor activity rhythms in larvae and adult armyworms.     

Circadian activity rhythms in cockroaches. 3. The role of endocrine and neural factors

The control of circadian activity rhythms (diurnal rhythms) in insects has been suggested to result by periodic neuroendocrine secretions. More specifically, Harker ('56) claimed that the locomotor rhythm in the cockroach, Periplaneta americana, is timed by a secretory “clock” located in the subesophageal ganglion. Later experiments by Harker indicated that this “clock” function failed unless the retrocerebral organs were left intact; allatectomy was said (no evidence given) to abolish a rhythm. The procedure for demonstrating a “clock” function in the ganglion involved transplanting it from a rhythmic donor into the hemocoel of an arrhythmic host and observing that the host subsequently became rhythmic. This result (without explicit information about the phase of the rhythm) does not warrant the conclusion that the ganglion acts as a clock. Therefore, I have attempted to confirm and extend these important results. Employing techniques essentially identical to Harker's, and using the same species of roach, I have been unable to find any evidence to support the original claim: (1) in 20 test animals, implantation of ganglia from rhythmic donors failed to re-instate a rhythm, and (2) allatectomy (22 cases) or removal of the entire retrocerebral complex (20 cases) did not interfere with the rhythm. The results of another series of experiments show that the cockroach brain is involved in the control of the activity rhythm. When the brain is surgically bisected (mid-sagittal) through the pars intercerebralis, arrhythmic activity patterns are immediately evoked. These continue for many weeks, but in a few cases rhythms ultimately “regenerate”.     

Increased size due to larval royal jelly exposure does not affect circadian locomotor activity or climbing ability in adult female Drosophila melanogaster

The effects of royal jelly (RJ) appear to be conserved in Drosophila; flies exposed of RJ exhibit increased body size, similar to queen bees. However, in flies and bees, there is evidence that increased body size can lead to impairments to locomotor activity, while RJ may have anti-fatigue properties. Canton-S and Oregon-R Drosophila larvae were raised on media containing 0% or 20% pure RJ. Climbing assays were conducted to assess vertical locomotion. Circadian locomotion was observed using Drosophila Activity Monitors. CS, but not Or-R, raised on RJ were larger compared to controls. Flies exposed to RJ exhibited entrainment and free-running rhythms. The increased size due to RJ exposure in this study had no bearing on circadian locomotor activity or climbing. These results indicate that there is variation among physiological responses to RJ among different strains, but RJ was equally ineffective in affecting locomotor behavior no matter the physiological response.     

The Correlation Between Activity,Blood Pressure and Heart Rate in Freely Mobile Sprague-Dawley and Transgenic Rats

Systolic (SBP) and diastolic blood pressure (DBP), heart rate (HR) and locomotor activity have been measured at 1-min intervals for 24 h in Sprague-Dawley (N = 5) and for 2x24 h in transgenic hypertensive (N = 4) rats. The animals were freely mobile and entrained to a 12:12 LD cycle (lights on at 0700). The endogenous circadian component of the cardiovascular variables was removed from the raw data, and then correlations between activity and the residual component (raw data minus the endogenous component) of SBP, DBP and HR were calculated. This calculation was performed twice, in the mid-light and mid-dark phases. We have investigated if the mean size of the correlation coefficients depended on cardiovascular variable (SBP, DBP or HR), phase (D or L) or strain (Sprague-Dawley, SPD, or Transgenic, TG, rats). Nearly all correlations were positive and ANOVA's showed a significant effect of cardiovascular variable for both strains, with correlations for HR being significantly higher than those for SBP and DBP. The mean correlations in the SPD strain were significantly higher than in the TG strain for variables SBP and DBP, but not for HR. The correlations between activity and blood pressure were more marked for SPD rats in the light (inactive) than dark (active) phase. Both strains showed ultradian rhythms in all variables, particularly in the light phase. If the analysis was repeated using deviations of the cardiovascular variables from a 1-h moving average rather than the endogenous circadian component, then the results were very similar. The results are discussed in terms of the links between the rhythms of activity and cardiovascular variables, with particular reference to differences between the two strains.     

The Influence of the Athymic Mutation Nude on the Components of the Orcadian Rhythm of Activity in Mice

The mutation known as nude brings about the lack of a thymus gland in mice. This immuno-deficiency makes it possible to graft normally unaccepted, human cancerous tumors onto the mouse. Consequently, this animal is frequently used as a model for evaluating anti-cancer therapies. The effect of this mutation on biological rhythms constitutes a necessary step before using this model for cancer chronotherapy research. We evaluated the circadian and ultradian components of the restactivity cycle in the following strains of mice: CS7BL/6 with homozygous nu/nu, heterozygous nu/+, thymectomised +/+, and sham-operated +/+. The amount of activity was reduced in nu/nu as compared to the other groups. Nonetheless, neither the nude mutation nor thymectomy yielded any notable change in the circadian rhythm of activity.     

Circannual cycles in golden-mantled ground squirrels: fall and spring cold pulses

Summary Endogenous circannual rhythms of male golden-mantled ground squirrels (Spermophilus lateralis) exposed to low ambient temperature (6 °C) at various times were compared to the rhythms of a control group kept in a warm room (21 °C) throughout. A 20-week pulse of cold in the spring delayed the subsequent peak body weight and molt offset. However, in the second year after the spring cold pulse, a delay in the rhythm was evident only using peak weight as a phase marker. A 20-week cold pulse in the fall resulted in an earlier peak body weight and earlier onset of pigmented scrotum and descended testes, but there was no evidence from subsequent years that rhythms had been phase advanced. A third of the animals kept continuously at 6 °C remained with pigmented scrotum, descended testes and low body weight for more than a year. Circannual periodicities of animals that continued to display rhythms in the cold room were not significantly longer than those of controls in the warm room. The results suggest that low temperature in the fall can alter the expression of circannual rhythms without much affecting their phasing, while low temperature in the spring produces phase delays which last longer but have not been proved to be persistent.Abbreviation PRC phase response curve     

CIRCADIAN RHYTHMS OF AGITATION IN INSTITUTIONALIZED PATIENTS WITH ALZHEIMER'S DISEASE

Agitation is a common problem in institutionalized patientswith Alzheimer's disease (AD). “Sundowning,” or agitationthat occurs primarily in the evening, is estimated to occur in 10—25%of nursing home patients. The current study examined circadian patterns ofagitation in 85 patients with AD living in nursing homes in the San Diego,California, area. Agitation was assessed using behavioral ratings collectedevery 15 minutes over 3 days, and activity and light exposure data were collectedcontinuously using Actillume recorders. A five-parameter extension of thetraditional cosine function was used to describe the circadian rhythms. Themean acrophase for agitation was 14:38, although there was considerable variabilityin the agitation rhythms displayed by the patients. Agitation rhythms weremore robust than activity rhythms. Surprisingly, only 2 patients (2.4%) were“sundowners.”In general, patients were exposed to very low levelsof illumination, with higher illumination during the night being associatedwith less robust agitation rhythms with higher rhythm minima (i.e., some agitationpresent throughout the day and night). Seasonality was examined; however,there were no consistent seasonal patterns found. This is the largest studyto date to examine agitation rhythms using behavioral observations over multiple24h periods. The results suggest that, although sundowning is uncommon, agitationappears to have a strong circadian component in most patients that is relatedto light exposure, sleep, and medication use. Further research into the understandingof agitation rhythms is needed to examine the potential effects of interventionstargeting sleep and circadian rhythms. (ChronobiologyInternational, 17(3), 405–418, 2000)     

Microbial hydroxylation. I. Hydroxylation of aniline byAspergillus alliaceus,A. albertensis andA. terreus

Summary Six hundred and seventy microorganisms were screened for the ability to perform stereoselective aromatic hydroxylation reactions of industrial significance, using aniline as a model substrate. TLC and HPLC analyses with diode array detection were used to identify and characterize hydroxylase activities. Of 79 cultures belonging to the speciesAspergillus alliaceus, A. albertensis, andA. terreus, 26 strains produced 2-aminophenol. Thirty strains were able to hydroxylate aniline in thepara position. Five strains ofA. terreus produced an unidentified phenolic compound in high yield.The mention of firm names or trade products does not imply that they are endorsed or recommended by the US Department of Agriculture over other firms or similar products not mentioned.     

Rhythmic gene expression in a purple photosynthetic bacterium, Rhodobacter sphaeroides

Circadian rhythms are known to exist in all groups of eukaryotic organisms as well as oxygenic photosynthetic bacteria, cyanobacteria. However, little information is available regarding the existence of rhythmic behaviors in prokaryotes other than cyanobacteria. Here we report biological rhythms of gene expression in a purple bacterium Rhodobacter sphaeroides by using a luciferase reporter gene system. Self-bioluminescent strains of Rb. sphaeroides were constructed, which produced a bacterial luciferase and its substrate, a long chain fatty aldehyde, to sustain the luminescence reaction. After being subjected to a temperature or light entrainment regime, the reporter strains with the luciferase genes driven by an upstream endogenous promoter expressed self-sustained rhythmicity in the constant free-running period. The rhythms were controlled by oxygen and exhibited a circadian period of 20.5 h under aerobic conditions and an ultradian period of 10.6-12.7 h under anaerobic conditions. The data suggest a novel endogenous oscillation mechanism in purple photosynthetic bacteria. Elucidation of the clock-like behavior in purple bacteria has implications in understanding the origin and evolution of circadian rhythms.     

Irregular 24-hour activity rhythms and the metabolic syndrome in older adults

Circadian rhythms – near 24?h intrinsic biological rhythms – modulate many aspects of human physiology and hence disruption of circadian rhythms may have an important impact on human health. Experimental work supports a potential link between irregular circadian rhythms and several key risk factors for cardiovascular disease including hypertension, obesity, diabetes and dyslipidemia, collectively termed the metabolic syndrome. While several epidemiological studies have demonstrated an association between shift-work and the components of the metabolic syndrome in working-age adults, there is a relative paucity of data concerning the impact of non-occupational circadian irregularity in older women and men. To address this question, we studied 7 days of actigraphic data from 1137 older woman and men participating in the Rush Memory and Aging Project, a community-based cohort study of the chronic conditions of aging. The regularity of activity rhythms was quantified using the nonparametric interdaily stability metric, and was related to the metabolic syndrome and its components obesity, hypertension, diabetes and dyslipidemia. More regular activity rhythms were associated with a lower odds of having the metabolic syndrome (OR?=?0.69, 95% CI?=?0.60–0.80, p?=?5.8?×?10^?7), being obese (OR?=?0.73, 95% CI?=?0.63–0.85, p?=?2.5?×?10^?5), diabetic (OR?=?0.76, 95% CI?=?0.65–0.90, p?=?9.3?×?10^?4), hypertensive (OR?=?0.78, 95% CI?=?0.66–0.91, p?=?2.0?×?10^?3) or dyslipidemic (OR?=?0.82, 95% CI?=?0.72–0.92, p?=?1.2?×?10^?3). These associations were independent of differences in objectively measured total daily physical activity or rest, and were not accounted for by prevalent coronary artery disease, stroke or peripheral artery disease. Moreover, more regular activity rhythms were associated with lower odds of having cardiovascular disease (OR?=?0.83; 95% CI?=?0.73–0.95, p?=?5.7?×?10^?3), an effect that was statistically mediated by the metabolic syndrome. We conclude that irregular activity rhythms are associated with several key components of the metabolic syndrome in older community-dwelling adults, and that the metabolic syndrome statistically partially mediates the association between activity rhythms and prevalent cardiovascular disease. Although additional longitudinal and experimental studies are needed to conclusively delineate the causal relationships underlying these associations, these findings are consistent with preclinical data, and add further support for investigations of the irregularity of activity rhythms as a potential therapeutic target to decrease the burden of cardiovascular disease in older adults.     

Selection of Portuguese Rhizobium leguminosarum bv. trifolii strains for production of legume inoculants

From several native clover species, growing in six different soil types, 170 Rhizobium leguminosarum biovar trifolii strains were isolated, covering the central and southern regions of Portugal. The effectiveness of the strains varied from ineffective to highly effective on T. subterraneum cv. Clare and on T. fragiferum cv. Palestine, with a predominance of medium and high effectiveness on both host plants. The effectiveness was not influenced by provenence (soil or plant), except for the strains from the rankers soils and for the strains isolated from T. pratense, that were ineffective or medium effective on T. subterraneum.Selected strains were evaluated for effectiveness on T. subterraneum cv. Clare, using the commercial strain TA1 as reference. Several of the isolated strains were more effective than TA1, indicating that local strains may be used to produce better inoculants.     

Variation in host specificity and gene content in strains from genetically isolated lineages of the ectomycorrhizal fungus Paxillus involutus s. lat.

Ectomycorrhizal fungi are known to vary in host range. Some fungi can enter into symbiosis with multiple plant species, while others have restricted host ranges. The aim of this study was to examine variation in host specificity among strains from the basidiomycete Paxillus involutus s. lat. Recent studies have shown that this fungus consists of at least four genetically isolated lineages, phylogenetic species (PS) I (which corresponds to the morphological species Paxillus obscurosporus), PS II (P. involutus s. str.), PS III (Paxillus validus), and PS IV (not yet supported by any reference material). Thirty-five Paxillus strains of PS I to IV were examined in microcosms for their capacity to infect birch (Betula pendula) and spruce (Picea abies). Seventeen strains were compatible and formed mycorrhizae with both tree species. Seven strains were incompatible with both birch and spruce. The gene content in three pairs of incompatible and compatible strains PS I, II, and III were compared using microarray-based comparative genomic hybridizations. Of 4,113 P. involutus gene representatives analyzed, 390 varied in copy numbers in at least one of the three pairwise comparisons. Only three reporters showed significant changes in all three pairwise comparisons, and none of these were changed in a similar way in three comparisons. Our data indicate that changes in host range have occurred frequently and independently among strains in P. obscurosporus, P. involutus s. str., and P. validus. No evidence was obtained demonstrating that these changes have been associated with the gain or loss of similar genes in these three species.     

Feature identification in circadian rhythms of mice strains using in vivo information

The objective of this work was to identify strain-specific characteristics from real-time measurements of circadian rhythms of two inbred mouse strains. In particular, heart rate, temperature, and activity data collected from A/J and C57BL/6J (B6) mice using telemetry are analyzed. The influence of activity on heart rate and temperature is minimized by correlation analysis followed by regression analysis. The correlation analysis is used to determine the length of the activity data filter that results in the best correlation between activity data and heart rate or temperature. After the activity data are filtered, they are used in regression analysis. The temperature and heart rate rhythms obtained as the intercepts of the regression analysis are interpreted as the zero-activity rhythms and consequently are good estimates of the circadian rhythms. The circadian temperature rhythms for the B6 mice follow a smoother cosine-like time waveform, whereas those for the A/J mice follow a more square-wave-like waveform. To quantify the difference between these two temperature rhythms, a feature based on Fourier analysis of the time-series data is used. Detrended fluctuation analysis is used to identify features in the heart rate rhythms. The results of this work show that the features for the circadian temperature and heart rate rhythms can be used as distinguishing characteristics of the A/J and B6 strains. This work provides the foundation for future studies directed at investigating the influence of chromosomal substitutions on the regulation of circadian rhythms in these two strains.     

The effects of different light–dark cycles on the metabolism of the diazotrophic,unicellular cyanobacteria Cyanothece sp. ATCC 51142, and Cyanothecesp. PCC 7822

The diazotrophic unicellular cyanobacterium Cyanothece sp. ATCC 51142 demonstrates circadian patterns in nitrogenase activity, H₂ production and glycogen storage when grown under nitrogen‐fixing, 12:12 light:dark (L:D) conditions. In this study, we grew Cyanothece sp. ATCC 51142, and another strain in this genus, Cyanothece sp. PCC 7822, under long‐day (16:8 L:D) and short‐day (8:16 L:D) nitrogen‐fixing conditions to determine if they continued to display circadian rhythms. Both strains demonstrated similar circadian patterns for all three metabolic parameters when grown under long‐day conditions. However, the strains responded differently to short‐day growth conditions. Cyanothece sp. ATCC 51142 retained reasonable circadian patterns under 8:16 L:D conditions, whereas Cyanothece sp. PCC 7822 had quite damped patterns without a clear circadian pattern. In particular, glycogen storage changed very little throughout the day and we ascribe this to the difference in the type of glycogen granules in Cyanothece sp. PCC 7822 which has small β‐granules, compared to the large, starch‐like granules in Cyanothece sp. ATCC 51142. The results suggested that both mechanistic and regulatory processes play a role in establishing the basis for these metabolic oscillations.     

Life in Zero Magnetic Field. V. E. coli Resistance to Antibiotics

Twenty‐six E. coli strains, isolated from human subjects, were tested for antibiotic drug resistance using the dilution of antibiotic solutions in agar culture medium. The bacterial strains were then exposed to zero magnetic field in a well‐controlled laboratory area, where a Helmholtz coil compensated the local geomagnetic field. The exposure time to the zero magnetic field was 6 days. The antibiotic drugs with antimicrobial large action spectra used to evaluate bacteria resistance were ampicillin, ceftazidime, tetracycline, ofloxacin, and kanamycin. The aqueous solutions of drug had dilutions of 0.25, 0.50, 4, 8, 16, 32, 64, and 128 µm/mL, respectively. Two types of microorganisms were detected: strains sensitive and strains nonsensitive to geomagnetic field compensation. We found that the magnetic‐sensitive strains represent about one‐third of the analyzed samples, statistical analysis emphasizing the general tendency of diminishing resistance against antibiotics.