Menstrual cycle and respiratory symptoms in a general Nordic-Baltic population.

RATIONALE: There is little knowledge of variations in respiratory symptoms during the menstrual cycle in a general population, and potential modifying factors are not investigated. OBJECTIVES: To investigate menstrual cycle variation in respiratory symptoms in a large general population, using chronobiology methodology, and stratifying by body mass index (BMI), smoking, and asthma status. METHODS: A total of 3,926 women with regular cycles less than or equal to 28 days and not taking exogenous sex hormones answered a postal questionnaire regarding the first day of their last menstruation and respiratory symptoms in the last 3 days. Moving 4-day means were computed to smooth uneven records of daily sampling; best-fitting 28-day composite cosine curves were applied to each time series to describe rhythmicity. MEASUREMENTS AND MAIN RESULTS: Significant rhythmic variations over the menstrual cycle were found in each symptom for all subjects and subgroups. Wheezing was higher on cycle Days 10-22, with a midcycle dip near the time of putative ovulation (approximately Days 14-16) in most subgroups. Shortness of breath was higher on days 7-21, with a dip just before midcycle in many subgroups. Cough was higher just after putative ovulation for subjects with asthma, BMI greater than or equal to 23 kg/m(2), and smokers, or just before ovulation and menses onset for low symptomatic subgroups. CONCLUSIONS: Respiratory symptoms varied significantly during the menstrual cycle and were most frequent from the midluteal to midfollicular stages, often with a dip near the time of ovulation. The patterns varied by BMI, smoking, and asthma status. These relations link respiratory symptoms with hormonal changes through the menstrual cycle and imply a potential for individualized chronotherapy for respiratory diseases.

Circadian variations in clock gene expression of human bone marrow CD34+ cells.

Time-dependent variations in clock gene expression have recently been observed in mouse hematopoietic cells, but the activity of these genes in human bone marrow (BM) has so far not been investigated. Since such data can be of considerable clinical interest for monitoring the dynamics in stem/progenitor cells, the authors have studied mRNA expression of the clock genes hPer1 , hPer2, hCry1, hCry2, hBmal1, hRev-erb alpha, and hClock in human hematopoietic CD34-positive (CD34( +)) cells. CD34(+) cells were isolated from the BM samples obtained from 10 healthy men at 6 times over 24 h. In addition, clock gene mRNA expression was analyzed in the whole BM in 3 subjects. Rhythms in serum cortisol, growth hormone, testosterone, and leukocyte counts documented that subjects exhibited standardized circadian patterns. All 7 clock genes were expressed both in CD34(+) cells and the whole BM, with some differences in magnitude between the 2 cell populations. A clear circadian rhythm was shown for hPer1, hPer2, and hCry2 expression in CD34(+) cells and for hPer1 in the whole BM, with maxima from early morning to midday. Similar to mouse hematopoietic cells, h Bmal1 was not oscillating rhythmically. The study demonstrates that clock gene expression in human BM stem/progenitor cells may be developmentally regulated, with strong or weaker circadian profiles as compared to those reported in other mature tissues.

Chronobiological analysis of circadian patterns in transcription of seven key clock genes in six peripheral tissues in mice.

The molecular clock machinery in mammals consists of a number of clock genes (CGs) and their resultant proteins that form interlocking transcription-translation feedback loops. These loops generate and maintain the 24 h mRNA and protein oscillations and consequential biological and physiological rhythms. To understand whether peripheral oscillators share similarly-timed clock machinery, the temporal expression patterns of the seven recognized key CGs (mPer1, mPer2, mCry1, mCry2, mRev-erb alpha, mClock, and mBmal1) were examined simultaneously in six peripheral tissues in mice every 4 h for 24 h in synchronized light-dark conditions using real time PCR assays. Time series were analyzed for time-effect by ANOVA and for rhythm characteristics by the single cosinor fitting procedure. The expression levels of most CGs were comparable in liver, kidney, and spleen, but mBmal1 and mCry1 were more abundant in the thymus, and mPer1, mCry1, and mCry2 were more abundant in the testis. In addition, mCry2 was dramatically lower in the kidney, spleen, and thymus; mPer2 was significantly lower in the spleen, testis, and thymus; and all of the genes tested were strikingly less abundant in peripheral blood. A significant 24 h rhythmic component was found for each CG in the liver and kidney and for some CGs in other tissues. Of note, a 12 h ultradian rhythmic component was also found in mRNA expression for some CGs in several of the tissues and was the only significant oscillation observed for CGs in the testis. Ultradian oscillations were also observed for mPer1 in the testis (8 h) and thymus (12 h and 8 h) in a second study where mice were sampled every 2 h. The present results suggest that the functioning of the molecular circadian clock may be modified to some extent between peripheral tissues, as denoted by differences in amplitude and phasing, and operates differently or is less developed in tissues containing differentiating cells (i.e., testis and thymus), as denoted by the presence of ultradian patterns resulting in two or more peaks within 24 h.

Circadian expression of clock genes in purified hematopoietic stem cells is developmentally regulated in mouse bone marrow.

OBJECTIVE: Clock genes are known to mediate circadian rhythms in the central nervous system and peripheral organs. Although they are expressed in mouse hematopoietic progenitor and stem cells, it is unknown if they are related to circadian rhythms in these cells. We therefore investigated the 24-hour patterns in the activity of several clock genes in the bone marrow (BM) side population (SP) primitive stem cells, and compared these 24-hour patterns to clock gene variations in the whole BM and liver. METHODS: Cells were obtained from 84 B6D2F(1) mice in three replicate experiments on the second day after release into constant darkness from a standardizing light-dark schedule. mRNA expression of clock genes was measured with quantitative reverse transcriptase polymerase chain reaction. RESULTS: mPer2 displayed circadian rhythms in SP cells, whole BM, and liver cells. mPer1 and mRev-erb alpha showed a circadian rhythm in whole BM and liver, but not SP cells. mBmal1 was not expressed rhythmically in SP cells, nor in the whole BM, contrary to rhythms observed in the liver. CONCLUSIONS: With the exception of mPer2, most clock genes studied in primitive hematopoietic SP stem cells were not oscillating in a fully organized circadian manner, which is similar to immature cells in rapidly proliferating organs, such as the testis and thymus. These findings indicate that circadian clock gene expression variations in BM are developmentally regulated.

Season, sun, sex, and cervical cancer.

INTRODUCTION: Sunlight's UV B component, a known cellular immunosupressant, carcinogen, and activator of viral infections, is generally seasonally available. Venereal human papillomavirus (HPV) transmission, at least in part, causes cervical cancer. We have previously inspected the monthly rates of venereal HPV infection and sunlight fluency in Southern Holland over 16 consecutive years. Both peak in August with at least 2-fold seasonality. The amount of available sunlight and the rate of Papanicolaou (Pap) smear screen-detected HPV are positively correlated. We now investigate whether premalignant and malignant cervical epithelial changes are also seasonal and related to seasonal sunlight fluency. METHODS: We have studied >900,000 consecutive, serially independent, interpretable screening Pap smears obtained by a single cervical cancer screening laboratory in Leiden, Holland, during a continuous 16-year span from 1983 through 1998. The average monthly rates of premalignant and malignant epithelial change were inspected and the annual patterns contrasted to the annual pattern of sunlight fluency at this global location and to monthly average HPV infection rate. Because HPV is venereally transmitted, Dutch seasonal sexual behavior was evaluated by assessment of the annual pattern of Dutch conception frequency as a competing cause for cervical cancer seasonality. RESULTS: (a) Twice as many premalignant and malignant epithelial changes were found among Pap smears obtained in the summer months, with an August peak concurrent with histopathologic evidence of HPV infection and sunlight fluency in Southern Holland. (b) Monthly sunlight fluency is correlated positively with both the monthly rates of Pap smear-detected cervical epithelial dysplasia and carcinomatous histopathology, as well as HPV. (c) Conception frequency, in this location, peaks in Spring not summer, and has a 4.8% annual amplitude. CONCLUSIONS: (a) Cervical epithelial HPV infection and HPV-induced cervical epithelial dysplasia and carcinomatous change may each be novel sun exposure risks and thereby behaviorably avoidable. (b) Because screening Pap smears uncover many abnormalities that resolve spontaneously (false positives), these data may argue for screening and follow-up Pap smear examinations in seasons other than summer in the Northern Hemisphere, to diminish the false-positive smear rate. Global data are available to confirm and further test each of these conclusions.

Measurement in saliva from neurotypical adults of biomarkers pertinent to autism spectrum disorders.

AIM: Measure biomarkers pertinent to autism in saliva from humans. MATERIALS & METHODS: At 7:30 PM (reading instructions) and 8:30 PM (hearing instructions), neurotypical adults (6 M, 6 F) each spat into tubes containing protease inhibitors. Cells were counted, samples aliquoted, frozen and thawed. Rationale was given for choice of biomarkers. ELISA: CD26, IL-12, carnitine, C4B, GSH, GSSG, MT-2, testosterone, IFN-γ. Mass spectrometry: cystine, glutamine, glutamic acid, GABA, serotonin. Electrochemiluminescentimmunoassay: cortisol. Radioimmunoassay: melatonin. RESULTS: Cells averaged 2.16 × 106/ml. M > F: CD-26, C4B, MT-2. Testosterone, cortisol. Glutamine, glutamic acid, IFN-γ, melatonin and GSSG were measurable. Remaining biomarkers were measured in <50% of samples. Concentrations were equal at both times. CONCLUSION: Saliva can be collected by literate individuals without added instruction. Ten biomarkers were measurable.

Chronomics: circadian and circaseptan timing of radiotherapy, drugs, calories, perhaps nutriceuticals and beyond.

We suggest a putative benefit from timing nutriceuticals (substances that are both nutrients and pharmaceuticals) such as antioxidants for preventive or curative health care, based on the proven merits of timing nutrients, drugs, and other treatments, as documented, i.a., in India. The necessity of timing melatonin, a major antioxidant, is noted. A protocol to extend the scope of chronoradiotherapy awaits testing. Imaging in time by mapping rhythms and broader time structures, chronomes, for earliest diagnoses, for example detection of vascular disease risk, is recommended. The study of rhythms and broader chronomes leads to a dynamic functional genomics, guided by imaging in time of free radicals and antioxidants, amongst many other variables.

Non-photic, non-thermic circadecadal solar cycle interaction with cardiovascular circannual and circasemiannual variation in heated air-conditioned habitat.

In order to re-examine the extent to which secular circulatory variation can be resolved into possibly accountable, if not yet predictable behavior, the 15-year record of blood pressure and heart rate of an adult male cardiologist (Y.W.) is reanalyzed. Gliding spectral windows with an interval of 4 years progressively displaced throughout the data series examine monthly means from August 1987 to August 2002. A circannual variation is only intermittent and appears to drift for years. A circasemiannual variation is consistent and prominent, yet only for a fraction of the record examined. By contrast to the circadian rhythm in blood pressure and heart rate, which is reliably detectable in most subjects in clinical health, the circannual variation in the human circulation is inconsistent and hence should be monitored, as in this case for its assessment as one goes, a conclusion that also applies to the circasemiannual variation, prominent for years but not detected thereafter. The results prompt the search for interactions between the photic, thermic and societal effects of the seasons that, if solely pertinent, should yield a consistent 1-year spectral component. That non-photic effects may also play an important role may point to modulation by solar cycle stage and solar cycle number, perhaps mediated by the solar wind. This conclusion is extended in proof to the role played by the most recently discovered transannual component in human blood pressure and heart rate, which is a probable further signature of the solar wind. The beating of the transannual and circannual components documented on another data series may contribute to the lack of a consistent 1-year synchronized circannual variation in this case and many others.

Blood pressure self-surveillance for health also reflects 1.3-year Richardson solar wind variation: spin-off from chronomics.

Self-experimentation concerns not only scientists, but also each individual for the sake of his/her chronobiologic health and science literacy, eventually to be acquired in primary and secondary education. Public education ensures that everybody who knows how to read or write can dispense with the service of a costly scribe. At all ages, public education can teach equally well how to find out whether one's blood pressure (BP) and heart rate (HR) responds to an increase in sodium intake with a rise, with no change or with a decrease in BP. This task and many others could become a matter of informed self-surveillance. Whenever there are inter-individual, sometimes opposite differences in response, government-sponsored trials on groups that do not consider such differences cannot solve what only the individual can do, at first by help from schools. Eventually special institutions may be designed for chronomics, the monitoring, interpretation and archivization of chronomes (time structures; from chronos = time and nomos = rule) of biological variables, also charged with a demographic analyzing and reporting system. Each individual's properly coded record, to guard confidentiality, becomes part of a promptly accessible database for one's own needs and for society's requirements. What individuals and small groups started as chronobiology, what is immediately available on back burners, as a service by an international project on the biosphere and the cosmos (BIOCOS) (corne001@umn.edu) could become a public system of planned surveillance archivization of one's rhythms from womb to tomb. Alterations of a rhythm's amplitude or acrophase or of a deterministic or other chaotic endpoint, such as a correlation dimension and approximate entropy, or of a standard deviation, among a multitude of other endpoints, can signal (in the otherwise neglected normal range) reversible risk elevations. If these elevated risks are detected and prompt the institution of countermeasures, such prehabilitation can save the cost of rehabilitation or of long-term care after morbid events; suffering also can be prevented such as that by those who are unlucky enough to helplessly survive a massive brain, heart or societal "stroke". As an equally important dividend, science gains in basic and applied terms, as illustrated herein by the demonstration of a trans-year, an approximately 1.3 to 1.6-year, heretofore unknown component of the human BP and HR spectrum, beating with the circannual component and characterizing the same data. Chronomically interpreted self-monitoring is a civic duty for both one's health and everybody's science.

Circasemiannual chronomics: half-yearly biospheric changes in their own right and as a circannual waveform.

Geomagnetic activity has a strong half-yearly but no precise yearly component in its spectrum, as Armin Grafe suggested nearly half a century ago. We have postulated elsewhere that non-photic cycles such as those in geomagnetics may have signatures in the biosphere and vice versa that biological rhythms have likely counterparts in the physical environment. Accordingly, we document phenomena characterized by a prominent about half-yearly variation, re-analyzed to constitute the start of a transdisciplinary chronomic (time structural) map, aligning these conditions with a half-yearly cycle in the geomagnetic index Kp. At least some biospheric phenomena fitted concomitantly with 1- and 0.5-year cosine curves exhibit an amplitude (A) ratio of A(0.5-year)/A(1-year) larger than unity. Methodologically, it is pertinent that even if data were read off published graphs, the resulting analyses were practically the same as those in the original data received subsequently. The main point is a circasemiannual pattern in status epilepticus, in several morbid oral conditions, in the cell density of vasopressin-containing neurons in the human suprachiasmatic nuclei (SCN), in circulating melatonin at middle latitudes at night during years of minimal solar activity or around noon at high latitudes, and in an unusual circasemiannual aspect of a birth-month-dependence of human longevity. Others have asked whether annual rhythms in human reproduction are biological, sociological or both. We show some other possibilities herein, involving the physical environment, hardly to be neglected in the case of open systems. As to almost certainly multifactorial circasemiannual rhythms, geomagnetics may also be a signal, a proxy or a putative, at least partial mechanism. Geomagnetic activity is related in its turn to solar and galactic activity, and may be a marker for other cyclic events that affect the biosphere. The similarity of cycle lengths in itself can only be a hint prompting the search for causal relations.

System times and time horizons for biospheric near-matches of primarily non-photic environmental cycles.

System time is defined as the duration (T) and sampling density (deltat) of a given study. A time horizon is obtained by combining analytical results on endpoints of time structures (chronomes), usually on control groups, from accumulating diverse studies carried out with drastically different values of T and deltat. These two design considerations depend upon and are limited by resources. The desideratum of a small At e.g., for studies of chaotic endpoints, limits T, e.g., has thus far precluded a continuous mapping of decadal cycles in endpoints such as the approximate entropy or the correlation dimension. Time horizons are being documented for an increasing number of variables that undergo cycles with infra-annual frequencies - with one cycle in about one, two and/or five decades - in the biosphere as in the environment. These infra-annuals, in and around us, modulate ultra-annuals, including circadians, as well as about 7-day (circaseptan) and about half-yearly (circasemiannual) cycles, all prominent, e.g., in geophysical and biospheric specta. Neglect of infra-annual chronomics can lead to blunders. A statistically highly significant decrease in the excretion of steroidal metabolites (or in heart rate variability) may be misinterpreted as premature aging if it lasts for several years, and may be unnecessarily acted upon, e.g., by drugs. The decrease, however, may only be transient, occurring only during a given stage of a circadecadal cycle, and it may be followed for the next several years by an also spurious "rejuvenation", possibly misinterpreted as a drug effect. When recognized as the alternating stages of decadal cycles and assessed with an affordable deltat, infra-annuals of variables involved in major problems of our day, including homicides and wars, may lead us to manipulable internal or external mechanisms, and thus, eventually, possibly to countermeasures to crime and terror as well as to the optimization of aims such as spirituality.

Chronoastrobiology: proposal, nine conferences, heliogeomagnetics, transyears, near-weeks, near-decades, phylogenetic and ontogenetic memories.

"Chronoastrobiology: are we at the threshold of a new science? Is there a critical mass for scientific research?" A simple photograph of the planet earth from outer space was one of the greatest contributions of space exploration. It drove home in a glance that human survival depends upon the wobbly dynamics in a thin and fragile skin of water and gas that covers a small globe in a mostly cold and vast universe. This image raised the stakes in understanding our place in that universe, in finding out where we came from and in choosing a path for survival. Since that landmark photograph was taken, new astronomical and biomedical information and growing computer power have been revealing that organic life, including human life, is and has been connected to invisible (non-photic) forces, in that vast universe in some surprising ways. Every cell in our body is bathed in an external and internal environment of fluctuating magnetism. It is becoming clear that the fluctuations are primarily caused by an intimate and systematic interplay between forces within the bowels of the earth--which the great physician and father of magnetism William Gilbert called a 'small magnet'--and the thermonuclear turbulence within the sun, an enormously larger magnet than the earth, acting upon organisms, which are minuscule magnets. It follows and is also increasingly apparent that these external fluctuations in magnetic fields can affect virtually every circuit in the biological machinery to a lesser or greater degree, depending both on the particular biological system and on the particular properties of the magnetic fluctuations. The development of high technology instruments and computer power, already used to visualize the human heart and brain, is furthermore making it obvious that there is a statistically predictable time structure to the fluctuations in the sun's thermonuclear turbulence and thus to its magnetic interactions with the earth's own magnetic field and hence a time structure to the magnetic fields in organisms. Likewise in humans, and in at least those other species that have been studied, computer power has enabled us to discover statistically defined endogenous physiological rhythms and further direct effects that are associated with these invisible geo- and heliomagnetic cycles. Thus, what once might have been dismissed as noise in both magnetic and physiological data does in fact have structure. And we may be at the threshold of understanding the biological and medical meaning and consequences of these patterns and biological-astronomical linkages as well. Structures in time are called chronomes; their mapping in us and around us is called chronomics. The scientific study of chronomes is chronobiology. And the scientific study of all aspects of biology related to the cosmos has been called astrobiology. Hence we may dub the new study of time structures in biology with regard to influences from cosmo- helio- and geomagnetic rhythms chronoastrobiology. It has, of course, been understood for centuries that the movements of the earth in relation to the sun produce seasonal and daily cycles in light energy and that these have had profound effects on the evolution of life. It is now emerging that rhythmic events generated from within the sun itself, as a large turbulent magnet in its own right, can have direct effects upon life on earth. Moreover, comparative studies of diverse species indicate that there have also been ancient evolutionary effects shaping the endogenous chronomic physiological characteristics of life. Thus the rhythms of the sun can affect us not only directly, but also indirectly through the chronomic patterns that solar magnetic rhythms have created within our physiology in the remote past. For example, we can document the direct exogenous effects of given specific solar wind events upon human blood pressure and heart rate. We also have evidence of endogenous internal rhythms in blood pressure and heart rate that are close to but not identical to the period length of rhythms in the solar wind. These were installed genetically by natural selection at some time in the distant geological past. This interpretive model of the data makes the prediction that the internal and external influences on heart rate and blood pressure can reinforce or cancel each other out at different times. A study of extensive clinical and physiological data shows that the interpretive model is robust and that internal and external effects are indeed augmentative at a statistically significant level. Chronoastrobiological studies are contributing to basic science--that is, our understanding is being expanded as we recognize heretofore unelaborated linkages of life to the complex dynamics of the sun, and even to heretofore unelaborated evolutionary phenomena. Once, one might have thought of solar storms as mere transient 'perturbations' to biology, with no lasting importance. Now we are on the brink of understanding that solar turbulences have played a role in shaping endogenous physiological chronomes. There is even documentation for correlations between solar magnetic cycles and psychological swings, eras of belligerence and of certain expressions of sacred or religious feelings. Chronoastrobiology can surely contribute to practical applications as well as to basic science. It can help develop refinements in our ability to live safely in outer space, where for example at the distance of the moon the magnetic influences of the sun will have an effect upon humans unshielded by the earth's native magnetic field. We should be better able to understand these influences as physiological and mechanical challenges, and to improve our estimations of the effects of exposure. Chronoastrobiology moreover holds great promise in broadening our perspectives and powers in medicine and public health right here upon the surface of the earth. Even the potential relevance of chronoastrobiology for practical environmental and agricultural challenges cannot be ruled out at this early stage in our understanding of the apparently ubiquitous effects of magnetism and hence perhaps of solar magnetism on life. The evidence already mentioned that fluctuations in solar magnetism can influence gross clinical phenomena such as rates of strokes and heart attacks, and related cardiovascular variables such as blood pressure and heart rate, should illustrate the point that the door is open to broad studies of clinical implications. The medical value of better understanding magnetic fluctuations as sources of variability in human physiology falls into several categories: 1) The design of improved analytical and experimental controls in medical research. Epidemiological analyses require that the multiple sources causing variability in physiological functions and clinical phenomena be identified and understood as thoroughly as possible, in order to estimate systematic alterations of any one variable. 2) Preventive medicine and the individual patients'care. There are no flat 'baselines', only reference chronomes. Magnetic fluctuations can be shown statistically to exacerbate health problems in some cases. The next step should be to determine whether vulnerable individuals can be identified by individual monitoring. Such vulnerable patients may then discover that they have the option to avoid circumstances associated with anxiety during solar storms, and/or pay special attention to their medication or other treatments. Prehabilitation by self-help can hopefully complement and eventually replace much costly rehabilitation. 3) Basic understanding of human physiological mechanisms. The chronomic organization of physiology implies a much more subtle dynamic integration of functions than is generally appreciated. All three categories of medical value in turn pertain to the challenges for space science of exploring and colonizing the solar system. The earth's native magnetic field acts like an enormous umbrella that offers considerable protection on the surface from harsh solar winds of charged particles and magnetic fluxes. The umbrella becomes weaker with distance from the earth and will offer little protection for humans, other animals, and plants in colonies on the surface of the moon or beyond. Thus it is important before more distant colonization is planned or implemented to better understand those magnetism-related biological- solar interactions that now can be studied conveniently on earth. (ABSTRACT TRUNCATED)

Rhythms in human bone marrow and blood cells.

In 24h studies of bone marrow (BM), circadian stage-dependent variations were demonstrated in the proliferative activity of BM cells from subsets of 35 healthy diurnally active men. On an average, the percentage of total BM cells in deoxyribonucleic acid (DNA) synthesis phase was 188% greater at midday than at midnight (circadian rhythm: p = 0.018; acrophase or peak time of 13: 16h). Patients with malignant disease (n = 15) and a normal cortisol circadian rhythm showed higher fractions of BM cells in S-phase at midday. Colony-forming units--granulocyte/macrophage (CFU-GM), an indicator of myeloid progenitor cells, showed the same circadian variation as DNA S-phase (average range of change or ROC = 136%; circadian rhythm: p < 0.001; acrophase of 12:09h). Deoxyribonucleic acid S-phase and CFU-GM in BM both showed a circannual rhythm (p = 0.015 and 0.008) with an identical acrophase of August 12. The daily peak in BM glutathione content, a tripeptide involved in cellular defense against cytotoxic damage, preceded BM proliferative peaks by 4-5 h (ROC = 31-90%; circadian rhythm: p = 0.05; acrophase of 08:30h). Myeloid (ROC = 57%; circadian rhythm: p = 0.056; acrophase at 08:40h) and erythroid (ROC = 26%; circadian rhythm: p = 0.01; acrophase of 13:01h) precursor cells were positively correlated (r = 0.41; p < 0.001), indicating a circadian temporal relationship and equal influence on S-phase of total BM cells. Yield of positive selected CD34+ progenitor stem cells also showed significant circadian variation (ROC = 595%; circadian rhythm: p = 0.02; acrophase of 12:40h). Thus, the temporal synchrony in cell cycling renders BM cells more sensitive at specific times to hematopoietic growth factors and cell cycle-specific cytotoxic drugs. Moreover, proper timing of BM harvesting may improve progenitor cell yield. When using marker rhythms in the blood to allow for individualized timing of BM procedures, the times of low values in white blood corpuscles, neutrophils, and lymphocytes and high values in cortisol were predictive of the times of highest BM erythroid, myeloid, and total S-phase numbers occurring in the following 12 h.

Circadian response of annual weeds to glyphosate and glufosinate.

Five field experiments were conducted in 1998 and 1999 in Minnesota to examine the influence of time of day efficacy of glyphosate [N-(phosphonomethyl)glycine] and glufosinate [2-amino-4-(hydroxymethyl-phosphinyl)butanoic acid] applications on the control of annual weeds. Each experiment was designed to be a randomized complete block with four replications using plot sizes of 3 x 9 m. Glyphosate and glufosinate were applied at rates of 0.421 kg ae/ha and 0.292 kg ai/ha, respectively, with and without an additional adjuvant that consisted of 20% nonionic surfactant and 80% ammonium sulfate. All treatments were applied with water at 94 L/ha. Times of day for the application of herbicide were 06:00h, 09:00h, 12:00h, 15:00h, 18:00h, 21:00h, and 24:00h. Efficacy was evaluated 14 d after application by visual ratings. At 14 d, a circadian response to each herbicide was found, with greatest annual weed control observed with an application occurring between 09:00h and 18:00h and significantly less weed control observed with an application at 06:00h, 21:00h, or 24:00h. The addition of an adjuvant to both herbicides increased overall efficacy, but did not overcome the rhythmic time of day effect. Results of the multiple regression analysis showed that after environmental temperature, time of day was the second most important predictor of percent weed kill. Thus, circadian timing of herbicide application significantly influenced weed control with both glyphosate and glufosinate.

Circadian response of annual weeds in a natural setting to high and low application rates of four herbicides with different modes of actions.

Four herbicides [glyphosate (GLYT), an amino acid synthesis inhibitor; glufosinate (GLUF), a glutamine synthetase inhibitor; fomesafen (FOME), a protoporphyrinogen oxidase inhibitor, and chlorimuron ethyl (CLIM), an acetolactate synthase inhibitor] were used to examine the influence of time of day of application on the control of a variety of annual broadleaf weeds in field studies conducted in Minnesota (five studies on GLYT and GLUF, three studies on FOME and CLIM). All herbicides were applied with an adjuvant at recommended high and low (half or quarter strength) rates every 3h between 06:00 and 24:00h local time. Visual ratings of percent weed control evaluated at 14d were analyzed by herbicide and application rate for each study and across studies for time-of-day effect by analysis of variance (ANOVA) and single cosinor. A circadian response to each herbicide was found, with greatest weed control observed between 09:00 and 18:00h. Increasing the herbicide application rate did not overcome the time-of-day effect (ANOVA: p < or = 0.008 for time-of-day effect for each herbicide and application rate). The least-squares fit of a 24h cosine was significant (p < or = 0.001) for each herbicide and application rate, with double amplitudes of 18-82% (units = % visual control) and estimated peaks (acrophases) near midday between 12:40 and 13:35h. Analysis of residuals obtained from multiple regression that included weed height, herbicide rate, temperature, and relative humidity as independent factors also found a significant time-effect by both ANOVA and cosinor for each herbicide and rate, with acrophases advancing significantly by 3 to 7h for GLYT and GLUF, but not for FOME or CLIM. These results suggest that the four herbicides, while belonging to different families with different modes of action, may reveal different peak times of efficacy when adjusting for environmental factors. Nonetheless, each displays similar circadian patterns when influenced by these factors under natural seasonal field conditions. The within-day rhythmic differences found in weed control are large enough to warrant consideration of the practical financial and environmental importance of the time-of-day that these and other herbicides are applied.

GIGANTEA and SPINDLY genes linked to the clock pathway that controls circadian characteristics of transpiration in Arabidopsis.

Several "clock" genes that regulate the circadian system in Arabidopsis thaliana have been identified. The GIGANTEA (GI) gene has been shown to participate in the circadian system that is linked to overt rhythms in gene expression, leaf movements, hypocotyl elongation, and photoperiodic control of flowering in Arabidopsis. During continuous light (LL), circadian expression patterns in gi-2 mutants show reduced amplitudes and altered period lengths when compared with controls. Rhythms in stomatal function, such as transpiration, have been shown to be endogenous and persist in constant lighting conditions. In order to test for a physiologic variable that might be affected by the circadian clock via the GI gene, we compared circadian characteristics of transpiration between three Arabidopsis mutants (gi-2, spy-4, spy-4/gi-2) and wild-type (WT) controls in synchronized (LD for 2.5d) and free-running (LL for 3d) conditions. Each genotype showed a significant circadian rhythm in LD at p < 0.001, with acrophases located near the middle of the daily 14h L-span, with average amplitudes for WT: 18.9%, gi-2: 16.1%, spy-4: 7.7%, and spy-4/gi-2: 5.3%. On the first day in LL, the circadian amplitude was dramatically reduced to 3.1% for gi-2 compared with WT (11.9%), while amplitudes for spy-4 (6.9%) and spy-4/gi-2 (5.7%) were not significantly changed from LD. The amplitude for gi-2 remained low during days 2 (4.2%) and 3 (2.1%) in LL, while it slowly dampened for the WT (8.6 and 6.6%). The amplitudes for spy-4 (6.6%) and spy-4/gi-2 (5.6%) on day 2 in LL were indistinguishable from the LD span, but finally dampened on day 3 in LL (1.9 and 2.3%, respectively). These data suggest that transpiration is a physiologic variable controlled by a circadian system that involves both the GI and SPY proteins.

Cross-spectrally coherent ~10.5- and 21-year biological and physical cycles, magnetic storms and myocardial infarctions*

Magnetic storms trigger myocardial infarctions with mechanisms relating to heart rate variability. Solar cycle-to-solar cycle differences and solar cycle stage dependence shown herein may resolve prior controversy and serve to advocate coordinated worldwide systematically aligned biological and physical monitoring. * This paper was originally invited by the historian-geophysicist Wilfried SCHRöDER of Bremen, Germany, for his biographical "Encounters," and is to serve as an update on the project on the BIOsphere and the Cosmos (BIOCOS) and its offspring ICEHRV (Dr. Kuniaki Otsuka's International Chronome Ecologic Study of Heart Rate Variability). It is intended for distribution at a NATO conference on space weather hazards, organized by Dr. Ioannis Daglis, June 18-29, 2000.

Transdisciplinary unifying implications of circadian findings in the 1950s.

Afew puzzles relating to a small fraction of my endeavors in the 1950s are summarized herein, with answers to a few questions of the Editor-in-Chief, to suggest that the rules of variability in time complement the rules of genetics as a biological variability in space. I advocate to replace truisms such as a relative constancy or homeostasis, that have served bioscience very well for very long. They were never intended, however, to lower a curtain of ignorance over everyday physiology. In raising these curtains, we unveil a range of dynamics, resolvable in the data collection and as-one-goes analysis by computers built into smaller and smaller devices, for a continued self-surveillance of the normal and for an individualized detection of the abnormal. The current medical art based on spotchecks interpreted by reference to a time-unqualified normal range can become a science of time series with tests relating to the individual in inferential statistical terms. This is already doable for the case of blood pressure, but eventually should become possible for many other variables interpreted today only based on the quicksand of clinical trials on groups. These ignore individual differences and hence the individual's needs. Chronomics (mapping time structures) with the major aim of quantifying normalcy by dynamic reference values for detecting earliest risk elevation, also yields the dividend of allowing molecular biology to focus on the normal as well as on the grossly abnormal.

The Chronobra identifies prevailing mammary vascularity as a candidate variable in breast cancer post-operative outcome prediction.

We previously described a menstrual heat cycle of the breast in four groups of women (healthy, family history of breast cancer, benign breast disease, 'cancer-associated') who wore a thermometric brassiere (Chronobra). We now ask if 'breast minus oral temperature', indicating 'breast-associated vascularity', could be associated with breast cancer cell vascular access around different aspects of the menstrual cycle rhythm and survival. Thirty-six pre-menopausal breast cancer patients (average age: 38.97 y) were enrolled consecutively over 15 y and followed for more than 22 y after surgery in order to compare survival and peri-operative vascularity. Each subject wore the Chronobra, which provides an internal bioassay of the vascularity of both breasts, including the operated breast, during 1 h each evening at home for one menstrual cycle, and collected saliva for "free" progesterone to confirm pre-menopausal status and ovulation. Sixty-five healthy age-matched pre-menopausal women served as controls. Both oral and breast temperatures revealed menstrual cycle oscillations, rising just before ovulation until menses onset. Breast-adjusted vascularity also showed menstrual cycle oscillations, with levels differing significantly between the 3 groups during the luteal phase only. At the end of the follow-up span, 18 post-operative breast cancer patients had died from "disseminated" breast cancer and 18 were alive and well. Median follow-up time was 22.6 y for survivors, 6.2 y for non-survivors, and 21.0 y for controls (3 died from diseases unrelated to breast cancer). Based on 'during luteal-phase breast-adjusted vascularity', breast cancer survivors (mean ± SD: -1.65 ± 0.23°C) were significantly hypo-vascular (i.e., -0.23°C cooler) compared with controls (-1.42 ± 0.09°C), while non-survivors (-1.25 ± 0.12°C) were highly significantly hyper-vascular compared with survivors (+0.41°C warmer) and controls (+0.23°C warmer). This suggests that in pre-menopausal breast cancer patients, peri-operative mammary vascularity could offer an outcome test of survival and biologically may be on the "final common pathway" of any tumor to metastatic risk and recurrence.

Determination of whole body circadian phase in lung cancer patients: melatonin vs. cortisol.

BACKGROUND: A quantifiable and reliable technique for the determination of body circadian phase applicable to non-laboratory studies would allow the evaluation of circadian dysregulation. In this study we evaluated feasible methodologies to individualize whole body circadian phase in lung cancer patients. METHODS: Cortisol and melatonin serum levels were measured in blood samples collected every 4 h for 24 h from eleven male controls and nine men suffering from non-small cell lung cancer. Circadian rhythmicity was evaluated and the 4-hourly fractional variations (FV) were calculated to evaluate the dynamics of the rise and fall in serum levels. RESULTS: Overall cortisol serum levels were higher in cancer patients (p<0.001), and lower for melatonin, but not significantly (p=0.261). Original serum levels of cortisol and melatonin each showed a prominent 24 h oscillation in both study groups, with highest values at night for melatonin and near awakening for cortisol. Using all data after normalization to percent of individual mean, ANOVA detected a significant time-effect (p<0.001) and cosinor analysis detected a significant 24 h rhythm (p<0.001) in each group. Overall fractional variation (FV) levels were lower for cortisol in cancer patients and higher for melatonin, but these differences were not significant. FV levels of cortisol and melatonin each showed a prominent 24 h oscillation in both study groups, with highest values prior to darkness onset for melatonin and near mid-dark for cortisol. ANOVA also detected a significant time-effect (p<0.002) and cosinor analysis detected a significant 24 h rhythm (p<0.001) for FV in each group, with maximal FV preceding maximal serum levels by ∼3 h for melatonin and ∼5 h for cortisol. CONCLUSIONS: A chronobiological evaluation of serum levels and fractional variations for cortisol and especially melatonin is a valuable methodology to define body circadian phase in lung cancer patients. It is possible to describe the complex process of hormone secretion with a methodology that allows the definition of both temporal characteristics and dynamic components. IMPACT: This kind of analysis might be useful in the study of hormone secretion(s) in cancer patients and other diseases and to guide therapeutic interventions. While lung cancer patients may have a negative prognostic value based upon stage and/or other hormonal aspects of their hypothalamus-pituitary-thyroid-adrenal axis function, patients that nevertheless maintain circadian rhythmicity in key body rhythm markers may still be eligible candidates for a chronotherapeutic approach of treatment(s).