Disrupted seasonal biology impacts health, food security and ecosystems.

The rhythm of life on earth is shaped by seasonal changes in the environment. Plants and animals show profound annual cycles in physiology, health, morphology, behaviour and demography in response to environmental cues. Seasonal biology impacts ecosystems and agriculture, with consequences for humans and biodiversity. Human populations show robust annual rhythms in health and well-being, and the birth month can have lasting effects that persist throughout life. This review emphasizes the need for a better understanding of seasonal biology against the backdrop of its rapidly progressing disruption through climate change, human lifestyles and other anthropogenic impact. Climate change is modifying annual rhythms to which numerous organisms have adapted, with potential consequences for industries relating to health, ecosystems and food security. Disconcertingly, human lifestyles under artificial conditions of eternal summer provide the most extreme example for disconnect from natural seasons, making humans vulnerable to increased morbidity and mortality. In this review, we introduce scenarios of seasonal disruption, highlight key aspects of seasonal biology and summarize from biomedical, anthropological, veterinary, agricultural and environmental perspectives the recent evidence for seasonal desynchronization between environmental factors and internal rhythms. Because annual rhythms are pervasive across biological systems, they provide a common framework for trans-disciplinary research.

Toxicity and clinical outcomes of paliperidone exposures reported to U.S. Poison Centers.

CONTEXT: Paliperidone is an atypical antipsychotic that was approved in the U.S. in 2006, and is also available in Canada, Australia, New Zealand, Europe, and Asia. Information regarding paliperidone overdoses is limited to case reports. Serious toxicity has yet to be reported. OBJECTIVE: To evaluate the toxicity of paliperidone exposures using a national poison center database. METHODS: A retrospective, observational case series of single-substance paliperidone cases reported to the National Poison Data System from 2007 to 2012 was conducted. Cases were evaluated for demographics, reason for exposure, clinical effects, treatments, disposition, and coded medical outcomes. For cases with major effects the text fields in poison center charts were evaluated to verify accuracy of coded outcome. The relationship between dose and severity of medical outcome was analyzed for acute exposure cases. RESULTS: There were 801 paliperidone cases that met inclusion criteria that included 592 persons of 13 years or greater, 67 children of 6-12 years, 140 children of less than 6 years, and 2 unknown ages. Most common reasons for exposure included: suicide attempt (39.6%), unintentional general (21.1%), therapeutic error (15.7%), and adverse drug reaction (11.9%). The most commonly observed clinical effects were drowsiness/lethargy (28.7%), tachycardia (23.3%), and dystonia (14.2%). Most patients were managed in the emergency department (40.3%) or were admitted to a health care facility (HCF) (42.7%). In 564 cases treated in a HCF, treatments included activated charcoal (25.7%), antihistamines (21.1%), and benzodiazepines (9.4%). Medical outcomes were no effect (35.0%), minor (30.8%), moderate (33.7%), and major effect (0.5%). There were no deaths. Of 491 acute exposures, dose was coded for 74.3% of exposures. There was a significant difference in the reported median dose between those with no effect (6 mg) and either minor effect (12 mg; p = 0.047) or moderate effect cases (12 mg; p = 0.020) in 91 children less than 6 years. CONCLUSIONS: The majority of patients experienced no or minor toxicity and were not admitted for medical care. Although a higher dose was associated with a more serious outcome in children less than 6 years, the data do not provide clear-cut triage guidelines.

Pregnancy modulates precursor cell proliferation in a murine model of focal demyelination.

In mice, pregnancy has been shown to have a beneficial effect on the endogenous repair of focal lysolecithin-induced CNS demyelinative lesions, enhancing the genesis of new oligodendrocytes and the degree of remyelination. To identify local cells undergoing mitosis in response to such lesions, we examined the time course of phospho-histone H3 (PH3) and myelin basic protein (MBP) expression by immunohistochemistry. After lysolecithin injection into the corpus callosum of virgin female mice, the number of dividing cells peaked about 48 h after injection and declined gradually to baseline by day 7; in pregnant mice, this initial peak was unchanged, but a new delayed peak on day 4 was induced. Colocalization data using PH3 and NG2 proteoglycan, or bromodeoxyuridine (BrdU) and oligodendrocyte transcription factor 1 (Olig1), suggested that about 75% of the proliferating cells on day 2, and about 40% of the cells on day 4, were likely of oligodendrocyte lineage; these differential percentages were of the same magnitude in both virgin and pregnant animals. Notably, the heightened proliferative response to focal lysolecithin injection during pregnancy was specific to gestational stage (early, but not late) and to lesion location (in the corpus callosum of the periventricular forebrain, but not in the caudal cerebellar peduncle of the hindbrain).

A MEMS-based high frequency x-ray chopper.

Time-resolved x-ray experiments require intensity modulation at high frequencies (advanced rotating choppers have nowadays reached the kHz range). We here demonstrate that a silicon microlever oscillating at 13 kHz with nanometric amplitude can be used as a high frequency x-ray chopper. We claim that using micro-and nanoelectromechanical systems (MEMS and NEMS), it will be possible to achieve higher frequencies in excess of hundreds of megahertz. Working at such a frequency can open a wealth of possibilities in chemistry, biology and physics time-resolved experiments.

[Cloning and characterization of a large metagenomic DNA fragment containing glycosyl-hydrolase genes].

The problem of search for and characterization of enzymes synthesized by non-cultivated microorganisms is presently being settled by creating metagenomic libraries. A 6000-clone library with the average size of its inserts amounting to 15 bp has been constructed on the basis of total DNA isolated from cow rumen microorganisms. As the result of the screening of the library on plates with different substrates, a clone was selected that efficiently hydrolyzed lichenan and carboxymethylcellulose. The clone contained the recombinant plasmid pBlue-13 bearing a 12071 bp.-long metagenomic fragment carrying ten open reading frames, two of them being identified as glycosyl hydrolase genes. No homology of the metagenomic DNA with any known microorganism genomes was revealed. The amino acid sequence, deduced on the basis of frame 4 and denoted by Xyl3A, bears resemblance with beta-xylosidases of glycosyl hydrolase Family 3. Frame 6 encodes polypeptide Cel5A homologous to cellulases of glycosyl hydrolase Family 5. The amino acid sequences deduced on the basis of seven out of ten open reading frames were homologous to proteins of microorganisms belonging to the Bacteroides sp. family, the bacteria inhabiting mammalian intestines.

X-ray pushing of a mechanical microswing.

We report here for the first time the combination of x-ray synchrotron light and a micro-electro-mechanical system (MEMS). We show how it is possible to modulate in real time a MEMS mass distribution to induce a nanometric and tunable mechanical oscillation. The quantitative experimental demonstration we present here uses periodic thermal dilatation of a Ge microcrystal attached to a Si microlever, induced by controlled absorption of an intensity modulated x-ray microbeam. The mechanism proposed can be envisaged either for the detection of small heat flux or for the actuation of a mechanical system.

On the chronobiology of cohabitation.

Social regulation of animal circadian rhythms may enable individuals in a population to temporally synchronize or segregate their activities within the community. Relatively little is known about the mechanisms for such interindividual temporal adaptations or how the circadian system might be involved. The literature suggests that actual prolonged cohabitation might lead to robust effects on the rhythmicity of cohoused individuals but that these effects are not easily reproduced by indirect or pulsatile social contacts. We have begun to study the conditions under which such cohabitation effects might be revealed in the laboratory, and we present and discuss initial data that cohousing pairs of golden hamsters can result in a persistent change in the free-running circadian period of one of the two hamsters of the pair. We believe that analyzing the societal level of temporal organization, and ultimately dissecting its underlying mechanisms, will enrich our understanding of the circadian clock and its role in establishing ecological communities.

Lack of toxicity from pediatric beta-blocker exposures.

The risk of toxicity in a child who is unintentionally exposed to a beta-blocking drug remains uncertain. The current study further defines this risk, particularly in the common scenario of ingestion of one or two tablets. A prospective cohort of 208 pediatric patients, 6 months to 6 years of age, reported to two regional poison centers serves as the study population. Data were collected with a standardized instrument during the care of each patient and for a minimum of 24 hours after exposure. No instances of serious toxicity typical of beta-blocker intoxication, such as 'shock-like' states, arrhythmias or seizures were observed in this series. Furthermore, there were no reported episodes of hypoglycemia, symptomatic bradycardia or bronchospasm. Nine instances of altered mental status or behavioral changes were reported. All appeared to be minor in nature. The most serious outcome was charcoal aspiration during gastrointestinal decontamination. This study adds to a growing body of evidence suggesting that exposure to one or two beta-blocker tablets places children at very little, if any, risk of toxicity.

Tumour size distribution in mammography screening.

Many mammography screening programmes have not been able to show higher percentages of small invasive cancers detected at subsequent screens than at initial screens. This has been a matter of serious concern as it contradicts the very theory of screening. Based on data from the county of Fyn, Denmark we evaluated the distribution based on point estimates, as well as on the entire tumour size distribution. The programme changed the amount of tumours less than 15 or 20 mm, but did not change the amount of tumours less than 10 mm. We evaluated the entire tumour size distribution and found that screen number was the only significant factor, implying that the number of screens changed the tumour size distribution. We recommend that the entire tumour size distribution is used to evaluate the ability of a programme to detect small breast cancers, instead of only point estimates as has previously been the practice.

Behavioral arousal blocks light-induced phase advances in locomotor rhythmicity but not light-induced Per1 and Fos expression in the hamster suprachiasmatic nucleus.

Both photic and nonphotic stimuli entrain circadian rhythms. Although the adaptive significance of nonphotic clock resetting is unknown, one possibility is that nonphotic cues modulate circadian responses to light. Results of studies on the interaction between photic and nonphotic stimuli support this idea. During the day, light blocks the effects of nonphotic stimuli on the phase of locomotor rhythms and on expression of clock genes in suprachiasmatic nucleus (SCN) neurons. At night, novelty-induced activity prior to and during exposure to light attenuates the phase-shifting response to that light, but the effects of this manipulation on clock gene expression are unknown. The present experiments explore the interaction between behavioral state and response to light at the molecular level. We show that confining hamsters to novel wheels immediately after a light pulse during the late subjective night attenuates light-induced phase advances of wheel-running rhythms and the transient effects on circadian period. In contrast to the striking effect of novelty-induced activity on behavioral responses to light, Fos protein and Per1 mRNA were robustly expressed in the SCN of all light-pulsed animals, regardless of behavioral treatment. Our results are inconsistent with the idea that light and nonphotic stimuli block each other's effects on phase shifts by inducing or attenuating transcription of Per1. Photic regulation of clock genes and spontaneous rhythmic expression of clock genes are probably mediated by different mechanisms.

Breast cancer incidence after the start of mammography screening in Denmark.

Mammography screening may lead to overdiagnosis of asymptomatic breast cancers, that would otherwise not have given rise to clinical symptoms. This aspect was studied in three regional screening programmes in Denmark, which started in Copenhagen municipality, Fyn county, and Frederiksberg municipality in 1991, 1993, and 1994, respectively. In these regions, we compared time trends in incidence of invasive breast cancer with the rest of Denmark. Since the number of clinical mammograms was relatively low, it was reasonable to assume that the breast cancer incidence outside the three screening regions represented the incidence of a population with low-intensity opportunistic screening. In Copenhagen and Fyn, a prevalence peak in incidence was seen during the first invitation round. During the subsequent invitation rounds, the incidence dropped to a level in line with the incidence expected without screening. The pattern was different in the small municipality of Frederiksberg, where the sensitivity was low during the first invitation round. Inclusion of screen-detected ductal carcinoma in situ cases did not change these results. The experiences from Copenhagen and Fyn show that organised mammography screening can operate without overdiagnosis of breast cancer.

Towards the reconstruction of central nervous system white matter using neural precursor cells.

Epidermal growth factor-responsive neural precursor cells were used as donor cells for transplantation into wild-type and myelin-deficient shiverer (shi) mice. The cells engrafted robustly within the CNS following intracerebroventricular and cisternal transplantation in neonatal mice. The cells adopted glial phenotypes, and some functioned as oligodendrocytes, producing myelin basic protein and morphologically normal internodal myelin sheaths. When individual shi mice received two transplants (on post-natal days 1 and 3), donor-derived cells disseminated widely and expressed myelin basic protein in central white matter tracts throughout the brain.

Randomized comparison of enoxaparin, a low-molecular-weight heparin, with unfractionated heparin adjunctive to recombinant tissue plasminogen activator thrombolysis and aspirin: second trial of Heparin and Aspirin Reperfusion Therapy (HART II).

BACKGROUND: Adjunctive unfractionated heparin (UFH) during thrombolytic therapy for acute myocardial infarction (AMI) promotes the speed and magnitude of coronary artery recanalization and reduces reocclusion. Low-molecular-weight heparins offer practical and potential pharmacological advantages over UFH in multiple applications but have not been systematically studied as adjuncts to fibrinolysis in AMI. METHODS AND RESULTS: Four hundred patients undergoing reperfusion therapy with an accelerated recombinant tissue plasminogen activator regimen and aspirin for AMI were randomly assigned to receive adjunctive therapy for at least 3 days with either enoxaparin or UFH. The study was designed to show noninferiority of enoxaparin versus UFH with regard to infarct-related artery patency. Ninety minutes after starting therapy, patency rates (thrombolysis in myocardial infarction [TIMI] flow grade 2 or 3) were 80.1% and 75.1% in the enoxaparin and UFH groups, respectively. Reocclusion at 5 to 7 days from TIMI grade 2 or 3 to TIMI 0 or 1 flow and TIMI grade 3 to TIMI 0 or 1 flow, respectively, occurred in 5.9% and 3.1% of the enoxaparin group versus 9.8% and 9.1% in the UFH group. Adverse events occurred with similar frequency in both treatment groups. CONCLUSIONS: Enoxaparin was at least as effective as UFH as an adjunct to thrombolysis, with a trend toward higher recanalization rates and less reocclusion at 5 to 7 days.

Encoding le quattro stagioni within the mammalian brain: photoperiodic orchestration through the suprachiasmatic nucleus.

Within the suprachiasmatic nucleus (SCN) is a pacemaker that not only drives circadian rhythmicity but also directs the circadian organization of photoperiodic (seasonal) timekeeping. Recent evidence using electrophysiological, molecular, and genetic tools now strongly supports this conclusion. Important questions remain regarding the SCN's precise role(s) in the brain's photoperiodic circuits, especially among different species, and the cellular and molecular mechanisms for its photoperiodic "memory." New data suggesting that SCN "clock" genes may also function as "calendar" genes are a first step toward understanding how a photoperiodic clock is built from cycling molecules.

Neural precursor cells form rudimentary tissue-like structures in a rotating-wall vessel bioreactor.

We have analyzed the biology of embryonic, epidermal growth factor-responsive murine neural precursor cells cultured in the high-aspect ratio vessel (HARV). Within 2-3 d of rotary-cell culture, such cells formed multiple, macroscopic, three-dimensional structures that were orders of magnitude larger than the cellular clusters ("neurospheres") formed by these cells in conventional stationary-flask cultures. Each HARV structure was composed of a multilayered cellular shell surrounding one or more central cavities that were bordered by pyknotic cell nuclei. Although the cells in the HARV structures were more pleomorphic than those in neurospheres, the structures did not appear to represent primitive neural tumors: the formation of HARV structures by precursor cells was not an irreversible phenotypic change, and the structures did not originate from the clonal expansion of single-progenitor cells; the growth rate and invasiveness of the cells in HARVs were less than those in flasks; and HARV-cultured cells did not form tumors after subcutaneous inoculation into the flanks of NOD-scid/scid mice. Immunohistochemical analysis suggested that HARV structures might be novel "prototissues" characterized by a crude, but organized, architecture, with a surface layer of immature proliferating cells (nestin- and proliferating cell nuclear antigen-positive) that enclosed strata of more differentiated cells (beta-tubulin III- and glial fibrillary acidic protein-positive) within. Rotary-cell culture may have significant implications for the eventual utility of neural precursors for clinical neurotransplantation.

Assembling a clock for all seasons: are there M and E oscillators in the genes?

The hypothesis is advanced that the circadian pacemaker in the mammalian suprachiasmatic nucleus (SCN) is composed at the molecular level of a nonredundant double complex of circadian genes (per1, cry1, and per2, cry2). Each one of these sets would be sufficient for the maintenance of endogenous rhythmicity and thus constitute an oscillator. Each would have slightly different temporal dynamics and light responses. The per1/cry1 oscillator is accelerated by light and decelerated by darkness and thereby tracks dawn when day length changes. The per2 /cry2 oscillator is decelerated by light and accelerated by darkness and thereby tracks dusk. These M (morning) and E (evening) oscillators would give rise to the SCN's neuronal activity in an M and an E component. Suppression of behavioral activity by SCN activity in nocturnal mammals would give rise to adaptive tuning of the endogenous behavioral program to day length. The proposition-which is a specification of Pittendrigh and Daan's E-M oscillator model-yields specific nonintuitive predictions amenable to experimental testing in animals with mutations of circadian genes.

Origin of the hook effect in extrinsic photoconductors.

The response of extrinsic photoconductors to a step change in incident photon flux has long been known to exhibit a sharp transient feature, particularly at higher signal levels, known as the hook effect. We demonstrate experimentally and theoretically that the hook effect can be due to reduced illumination adjacent to the injecting contact. This nonuniformity can be produced by the transverse illumination of the detector that is common for far-infrared Ge:Ga devices. The hook effect has been demonstrated to be either present or absent in the same Ge:Ga photoconductor, at comparable signal size, depending on the nature of the contact illumination. Numerical finite-difference calculations of the transient response support this explanation and produce features that replicate the experimental results.

Antiphase oscillation of the left and right suprachiasmatic nuclei.

An unusual property of the circadian timekeeping systems of animals is rhythm "splitting," in which a single daily period of physical activity (usually measured as wheel running) dissociates into two stably coupled components about 12 hours apart; this behavior has been ascribed to a clock composed of two circadian oscillators cycling in antiphase. We analyzed gene expression in the hypothalamic circadian clock, the suprachiasmatic nucleus (SCN), of behaviorally "split" hamsters housed in constant light. The results show that the two oscillators underlying the split condition correspond to the left and right sides of the bilaterally paired SCN.