Temperature compensation in an ultradian rhythm of tyrosine aminotransferase activity inEuglena gracilis Klebs

Summary Tyrosine aminotransferase activity ofEuglena oscillates with an ultradian period of approximately 4–5 h. The oscillation frequency in the time series was determined by cosine fitting. Experiments which were performed between 16 and 31.5°C revealed temperature compensation.     

Oral melatonin produces arrhythmia in sparrows

Summary House sparrows,Passer domesticus, exhibit circadian rhythms of perch-hopping behavior. The rhythm was abolished by ad libitum administration of melatonin in the drinking water.Support was provided to S. Binkley by NSF PCM 8314331.     

Gonyauline: A novel endogenous substance shortening the period of the circadian clock of a unicellular alga

Summary The circadian clock in the unicellular algaGonyaulax polyedra is accelerated by a substance in extracts from the cells themselves. The extracts have been fractionated using the circadian rhythm of bioluminescence as bioassay. The active substance, termed gonyauline, has been isolated and characterized as a novel low molecular weight cyclopropanecarboxylic acid (S-methyl-cis-2-(methylthio) cyclopropanecarboxylic acid). Synthetic gonyauline has a similar shortening effect on the period of the circadian clock.     

The Dead Sea — alive again

In the thirteen years of quantitative studies on the microbiology of the Dead Sea from 1980 onwards three distinct periods can be discerned. Mass development of the green unicellular algaDunaliella parva (up to 8,800 cells/ml) and red archaeobacteria (2×10⁷ cells/ml) was observed in 1980, following a dilution of the upper water layers by rain floods. This bloom disappeared at the end of 1982 as a result of a complete mixing of the water column. During the period 1983–1991 the lake was holomictic, and noDunaliella cells were observed. Viable bacteria were present during this period in very low numbers. Heavy rain floods during the winter of 1991–1992 caused a new stratification as the upper five meters of the water column became diluted to 70% of their normal salinity. In this upper water layerDunaliella reappeared (up to 3×10⁴ cells/ml at the beginning of May, rapidly declining to less than 40 cells/ml at the end of July), and a bloom of red archaeobacteria (3×10⁷ cells/ml) once more imparted a red coloration to the lake.     

Ultradian (between 5 and 70 minutes) respiratory (VO2, VCO2) rhythms of 4 small vertebrates used in biomedical research]

Four laboratory Vertebrate species with a predominant activity, either diurnal (Coturnix coturnix japonica), nocturnal (Rattus norvegicus, Mus musculus), or equivocal (Cavia porcellus), present, in usual housing conditions (temperature 20 degrees C, hygrometry 60-70%, acoustical background 70 dB, LD12:12 lighting L = 150 lx), food and water ad libitum, ultradian respiratory (VO2, VCO2) rhythms of short periods. The frequencies of these rhythms are independent of food intake and activity variations correlated to the lighting phase and regimen (LD, LL). The period (tau) of these ultradian respiratory rhythms is in isolated animals, between 5 and 70 min., according to the species studied. Grouping of several individuals of a same species, in a ventilated chamber, modifies the statistical distribution of the periods of the ultradian rhythms, but does not change the interspecies differences.     

Phosphate transport inClaviceps sp. strain SD-58

Summary Phosphate uptake inClaviceps sp. strain SD-58 was found to be linear for 20 min, proportional to cell density in mg/ml, energy dependent, and taking place against a concentration gradient with a K_m value of 45.45×10^–5 M. Osmotic shock treatment to the cell caused a reduction in phosphate uptake associated with the release of binding protein. Partial restoration of uptake was observed on incubation of osmotically shocked cells with shock fluid. The results are discussed with reference to the effect of phosphate on alkaloid synthesis inClaviceps sp. strain SD-58.     

Expression of multidrug resistance (mdr) gene(s) in primary lymphoid organs of chicken immune system during embryonic development

The presence of a multidrug resistance (MDR) related protein, P-170, in normal and pathological lymphoid cells has been described. The present report evaluates the expression of themdr 1 gene by using the reverse Polymerase Chain Reaction (PCR) on cells obtained from the thymus and bursa of chicken embryos starting from day 12 until hatching. Results show that the thymic cells are positive from day 12 to the end of the observation period. In contrast,mdr mRNA was detected in the bursa from day 14 to day 17 of embryonic life. Possible relationships between the expression ofmdr and the development of T and B lymphocytes are discussed.     

Sex differences in human circadian rhythms: Intrinsic periods and sleep fractions

Summary The period of freerunning circadian rhythms is significantly shorter and the fraction of sleep is significantly larger in human females than in males, as long as the rhythms run internally synchronized. The sex difference in the period could be a property either of the whole circadian system or of only one of the oscillators in a multi-oscillator system. The sex difference in the sleep fraction could be a fixed property of the sleep-wake rhythm or could depend on interactions in the multi-oscillator system. To investigate these questions, a sample of 33 long-term experiments, in which the rhythms ran internally synchronized in one section and internally desynchronized in another section, were analyzed. The periods of rhythms in rectal temperature were different in females and males during internal synchronization, but became identical during internal desynchronization. In contrast, sex differences in sleep-wake periods were more pronounced when the rhythms were desynchronized than when they were internally synchronized. This result provides evidence that the sex difference in periodicity is a property only of the sleep-wake rhythm; the intrinsic periods of temperature rhythms are identical in females and males, whereas those of sleep-wake rhythms are distinctly shorter in females than in males. In the state of internal synchronization, the joint period is a compromise between the intrinsic periods of the rhythms involved, and therefore it shows a small but significant sex difference. Moreover, the transition from internally synchronized to desynchronized rhythms is combined with a highly significant reduction in the sleep fraction, which is considerably greater in females than in males. These results suggest that the occurrence of internal desynchronization strongly affects the sleep-wake rhythm, and that the influence of rhythm disorders is considerably greater in females than in males.     

Genetic and photoperiodic control of an avian reproductive cycle

European blackcaps,Sylvia atricapilla, with one breeding season per year, have a single-peaked annual testes cycle. However, African conspecifics from the Cape Verde Islands with two breeding seasons per annum demonstrate a two-peaked cycle. Both population-specific cycles reflect differences in the respective endogenous circannual rhythms. Experimental hybridization of birds of the two populations resulted in an intermediate pattern of testes cycle, thus demonstrating that there are genetic components for some temporal aspects in an avian reproductive cycle. Another characteristic of the African birds, their extremely rapid juvenile development and early sexual maturity (at an age of 5–6 months) proved largely to be a photoperiodic (short-day) effect in birds hatched in autumn. The same effect could also be induced in European conspecifics exposed to correspondingly short day-lengths.     

Mixed life cycles in the transitional zone between voltinisms in the fall webworm,Hyphantria cunea

The fall webworm,Hyphantria cunea Drury, was introduced from North America into Japan at half a century ago. At present, the populations north of 36° N are bivoltine and those south of 36° N are trivoltine. In the present study, the life cycle in the transitional zone between voltinisms was clarified in Tsukuba (36.1° N). When fourth instar larvae of the second generation were collected in the field and maintained in the laboratory, the incidence of pupal diapause increased when the sampling date was delayed from 16 August to 5 September. The fact that some developed without diapause inidcates the occurrence of a trivoltine life cycle in this locality, where a bivoltine life cycle is predominant. The critical photoperiod for diapause induction at 20 and 25°C was 14 h 13 min and 14 h 10 min, respectively. The photoperiodic response may explain the rapid increase in diapause incidence in late summer. In laboratory-reared diapause pupae, the time and period of chilling (5°C) greatly influenced the time required for adult emergence at the final incubation temperature of 25°C. However, diapause pupae chilled for a sufficiently long period developed to adulthood rapidly, irrespective of the conditions before chilling. Therefore, the difference in timing of entering diapause between pupae of the second and third generations would not result in their temporal reproductive isolation in the following spring. It is concluded that the mixed voltinism in Tsukuba is not a result of a mixture of genetically distinct bivoltine and trivoltine populations, but a result of the phenotypic plasticity in a genetically rather homogeneous population.     

Optic lobe neurosecretory cells of an Indian spiderCryptophora sp. (Areinidae)

Summary In all the 3 pairs of optic lobes ofCryptophora sp. there are small groups of monopolar PF^– neurosecretory cells which take on a green colour by PF technique and red by Azan. Their function is unknown, but they may be involved with photoperiodically controlled activity rhythms.We wish to express our thanks to Prof. U.S. Srivastava for providing working facilities and valuable suggestions.     

Osmotic adaptation of moderately halophilic methanogenic Archaeobacteria,and detection of cytosolicN,N-dimethylglycine

Methanohalophilus mahii SLP andMethanohalophilus halophilus Z-7982, two closely-related, moderately halophilic, methylotrophic methanogens, were tested for their adaptation to saline conditions. They grew in a wider range of salinities than previously reported, in a defined medium with as little as 0.1 M NaCl, and with a high as 4.0 M NaCl forM. halophilus and 4.5 M NaCl forM. mahii. Fastest growth occurred with 1.5 M NaCl forM. mahii and 1.0 M NaCl forM. halophilus. M. mahii also grew in media in which NaCl was replaced by sucrose or KCl as osmolytes up to the osmolal equivalent of 2 and 2.5 M NaCl (these media contained other sodium salts totaling about 0.1 M Na⁺). In media with either sucrose of KCl replacing NaCl,M. mahii grew fastest at osmolalities approximately equiosmolal to 1 M NaCl.M. mahii not only grew well at a wide range of osmosities, it also tolerated rapid shifts in osmolality. Cells subjected to a rapid 10-fold hypertonic shift resumed growth without a prolonged lag. When cells were subjected to a rapid 10-fold hypotonic shift, 90% of cells lysed, but the remaineder continued to swell with little further lysis during the next 45 min. Surviving cells resumed growth.Methanohalophilus strains grown in defined medium had low cytosolic Na⁺ concentrations; K⁺ concentrations were as high as 0.35 M. Organic osmotica in the cytosol include glycine betaine and larger amounts of N,N-dimethylglycine.     

Ultradian and other rhythms in human respiration rate

Summary Twelve male subjects had their respiration rate (RR) measured at 3 min intervals for 6 h. Although there were substantial individual differences, most subjects demonstrated a 90±15 min rhythm of RR, with several subjects also showing 60±10 min and 30±3 min rhythms.     

Transfer of secondary metabolites from the spongesDysidea fragilis andPleraplysilla spinifera to the mantle dermal formations (MDFs) of the mudibranchHypserlodoris webbi

The opisthobranch molluscHypselodoris webbi is able to select, among its potential preys, sponges chemically rich in furanosesquiterpenoids. The sequestered secondary metabolites act as defensive allomones against predators and are accumulated in some dorsal glands (MDFs). This transfer from sponges to MDFs has been proven by maintainingH. webbi together with some selected sponges in an aquarium for a prolonged period.     

Photoperiodic regulation of mating behaviour in the linden bug,Pyrrhocoris apterus,is mediated by a brain inhibitory factor

Active inhibition of mating behaviour in a male insect is reported here for the first time. InPyrrhocoris apterus L. (Heteroptera), the most important inhibitory pathway runs from the pars intercerebralis (PI) of the brain and does not pass through the corpora allata. The inhibitory activity of the PI is promoted by short day conditions and suppressed by long days. As the effect of photoperiod is delayed, transfer procedures enabled us to record daily rhythms in mating behaviour during short days. While the extirpation of the PI results in a discrete phase shift of the long day rhythm, there is a much less significant phase shift after this operation during short days. Thus the PI has been shown to mediate the effect of photoperiod on both the inhibition and the rhythm of mating behaviour.     

Variation in regulation of aflatoxin biosynthesis among isolates ofAspergillus flavus

Two new phenotypes ofAspergillus flavus which exhibit novel patterns of aflatoxin production have been identified and characterized. In one of the new variants ofA. flavus, aflatoxin is made in the absence of carbohydrate and concomitantly with growth, without a lag period. A second variant did not produce aflatoxin in the presence or absence of carbohydrate. Chemical mutagenesis of this nonaflatoxigenic strain resulted in mutant strains which produced aflatoxin on carbohydrate-containing media. The aflatoxin production pattern observed in these mutants resembled the typical production scheme, with a lag period through log phase growth.     

Rhythms of enzymatic activity in maternal and umbilical cord blood

The 24-h activity patterns of variouns enzymes were determined in human serum, red blood cells and white blood cells of maternal and umbilical cord blood. Blood was drawn from the brachial vein of mothers and from the umbilical cord within ten minutes after delivery. Corresponding blood specimens were obtained from 83 spontaneous labors, occurring at different hours over a period of 60 days. For each variable (variable=activity of a specific enzyme in one of the blood components) the results were grouped according to delivery hour, forming a 24-h pattern which was analyzed to elucidate time dependency. Five out of six corresponding maternal and fetal variables were similar with regard to pattern and peak time. The activity rhythms of glyceraldehyde-3-phosphate dehydrogenase and glucose phosphate isomerase in red blood cells of mothers and fetuses possessed a significant bimodal pattern. The activity rhythms of the latter enzyme in white blood cells and sera exhibited a significant 24-h period. Hexosaminidase activity exhibited a distinct 24-h rhythm in maternal white blood cells, but no significant rhythm could be detected in the fetal white blood cells. The activity of hexosaminidase showed, identical 24-h patterns in maternal and cord serum when analyzed by best fit cosine, and no significant time-dependency when analyzed by ANOVA.     

Rapid effects of insulin on cyclic GMP location in an intact protozoan

We studied rapid changes in location of cyclic GMP inTetrahymena pyriformis. Insulin caused cGMP localization in cilia and near the plasma membrane (0.5–1 min). Later (1–5 min) cGMP localization was diffuse in cytoplasm with perinuclear accentuation. Inactive insulin analogs did not elicit these changes.     

Rhythms of enzymatic activity in maternal and umbilical cord blood.

The 24-h activity patterns of various enzymes were determined in human serum, red blood cells and white blood cells of maternal and umbilical cord blood. Blood was drawn from the brachial vein of mothers and from the umbilical cord within ten minutes after delivery. Corresponding blood specimens were obtained from 83 spontaneous labors, occurring at different hours over a period of 60 days. For each variable (variable = activity of a specific enzyme in one of the blood components) the results were grouped according to delivery hour, forming a 24-h pattern which was analyzed to elucidate time dependency. Five out of six corresponding maternal and fetal variables were similar with regard to pattern and peak time. The activity rhythms of glyceraldehyde-3-phosphate dehydrogenase and glucose phosphate isomerase in red blood cells of mothers and fetuses possessed a significant bimodal pattern. The activity rhythms of the latter enzyme in white blood cells and sera exhibited a significant 24-h period. Hexosaminidase activity exhibited a distinct 24-h rhythm in maternal white blood cells, but no significant rhythm could be detected in the fetal white blood cells. The activity of hexosaminidase showed, identical 24-h patterns in maternal and cord serum when analyzed by best fit cosine, and no significant time-dependency when analyzed by ANOVA.     

Effects of the light-dark cycle and scheduled feeding on behavioral and reproductive rhythms of the cyprinodont fish,Medaka,Oryzias latipes

Summary Medaka were maintained on a 168 light-dark cycle and fed once daily on one of 5 different feeding schedules. The daily rhythm of agonistic behavior rapidly entrained to the scheduled feeding time and maintained this entrainment during a 3-day starvation period. In contrast the daily rhythms of egg laying and courtship stayed entrained to the L:D cycle regardless of the feeding schedule. Thus, temporal integration of this fish with its daily environment can involve multistimuli which concurrently and differentially entrain externally expressed circadian systems.We thank Mark Goodrich and Steve Huber for fish maintenance and technical assistance; and Don Dovala for aid in developing the automatic feeder. The research was funded in part by NIH, AM 25191 and NIEHS, ES No. 01985.