Twist1 Is a TNF-Inducible Inhibitor of Clock Mediated Activation of Period Genes.

BACKGROUND: Activation of the immune system affects the circadian clock. Tumor necrosis factor (TNF) and Interleukin (IL)-1β inhibit the expression of clock genes including Period (Per) genes and the PAR-bZip clock-controlled gene D-site albumin promoter-binding protein (Dbp). These effects are due to cytokine-induced interference of E-box mediated transcription of clock genes. In the present study we have assessed the two E-box binding transcriptional regulators Twist1 and Twist2 for their role in cytokine induced inhibition of clock genes. METHODS: The expression of the clock genes Per1, Per2, Per3 and of Dbp was assessed in NIH-3T3 mouse fibroblasts and the mouse hippocampal neuronal cell line HT22. Cells were treated for 4h with TNF and IL-1β. The functional role of Twist1 and Twist2 was assessed by siRNAs against the Twist genes and by overexpression of TWIST proteins. In luciferase (luc) assays NIH-3T3 cells were transfected with reporter gene constructs, which contain a 3xPer1 E-box or a Dbp E-box. Quantitative chromatin immunoprecipitation (ChIP) was performed using antibodies to TWIST1 and CLOCK, and the E-box consensus sequences of Dbp (CATGTG) and Per1 E-box (CACGTG). RESULTS: We report here that siRNA against Twist1 protects NIH-3T3 cells and HT22 cells from down-regulation of Period and Dbp by TNF and IL-1β. Overexpression of Twist1, but not of Twist2, mimics the effect of the cytokines. TNF down-regulates the activation of Per1-3xE-box-luc, the effect being prevented by siRNA against Twist1. Overexpression of Twist1, but not of Twist2, inhibits Per1-3xE-box-luc or Dbp-E-Box-luc activity. ChIP experiments show TWIST1 induction by TNF to compete with CLOCK binding to the E-box of Period genes and Dbp. CONCLUSION: Twist1 plays a pivotal role in the TNF mediated suppression of E-box dependent transactivation of Period genes and Dbp. Thereby Twist1 may provide a link between the immune system and the circadian timing system.

In vivo imaging of the central and peripheral effects of sleep deprivation and suprachiasmatic nuclei lesion on PERIOD-2 protein in mice.

STUDY OBJECTIVES: That sleep deprivation increases the brain expression of various clock genes has been well documented. Based on these and other findings we hypothesized that clock genes not only underlie circadian rhythm generation but are also implicated in sleep homeostasis. However, long time lags have been reported between the changes in the clock gene messenger RNA levels and their encoded proteins. It is therefore crucial to establish whether also protein levels increase within the time frame known to activate a homeostatic sleep response. We report on the central and peripheral effects of sleep deprivation on PERIOD-2 (PER2) protein both in intact and suprachiasmatic nuclei-lesioned mice. DESIGN: In vivo and in situ PER2 imaging during baseline, sleep deprivation, and recovery. SETTINGS: Mouse sleep-recording facility. PARTICIPANTS: Per2::Luciferase knock-in mice. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Six-hour sleep deprivation increased PER2 not only in the brain but also in liver and kidney. Remarkably, the effects in the liver outlasted those observed in the brain. Within the brain the increase in PER2 concerned the cerebral cortex mainly, while leaving suprachiasmatic nuclei (SCN) levels unaffected. Against expectation, sleep deprivation did not increase PER2 in the brain of arrhythmic SCN-lesioned mice because of higher PER2 levels in baseline. In contrast, liver PER2 levels did increase in these mice similar to the sham and partially lesioned controls. CONCLUSIONS: Our results stress the importance of considering both sleep-wake dependent and circadian processes when quantifying clock-gene levels. Because sleep deprivation alters PERIOD-2 in the brain as well as in the periphery, it is tempting to speculate that clock genes constitute a common pathway mediating the shared and well-known adverse effects of both chronic sleep loss and disrupted circadian rhythmicity on metabolic health.

Soil quality assessment through a multi-approach analysis in soils of abandoned terraced land in NE Spain

The abandonment of agricultural land in mountainous areas has been an outstanding problem along the last century and has captured the attention of scientists, technicians and administrations, for the dramatic consequences sometimes occurred due to soil instability, steep slopes, rainfall regimes and wildfires. Hidromorfological and pedological alterations causing exceptional floods and accelerated erosion processes has therefore been studied, identifying the cause in the loss of landscape heterogeneity. Through the disappearance of agricultural works and drainage maintenance, slope stability has resulted severely affected. The mechanization of agriculture has caused the displacement of vines, olives and corks trees cultivation in terraced areas along the Mediterranean catchment towards more economically suitable areas. On the one hand, land use and management changes have implicated sociological changes as well, transforming areas inhabited by agricultural communities into deserted areas where the colonization of disorganized spontaneous vegetation has buried a valuable rural patrimony. On the other hand, lacking of planning and management of the abandoned areas has produced badlands and infertile soils due to wildfire and high erosion rates strongly degrading the whole ecosystems. In other cases, after land abandonment a process of soil regeneration has been recorded. Investigations have been conducted in a part of NE Spain where extended areas of terraced soils previously cultivated have been abandoned in the last century. The selected environments were semi-abandoned vineyards, semi-abandoned olive groves, abandoned stands of cork trees, abandoned stands of pine trees, scrubland of Cistaceaea, scrubland of Ericaceaea, and pasture. The research work was focused on the study of most relevant physical, chemical and biological soil properties, as well as runoff and erosion under soils with different plant cover to establish the abandonment effect on soil quality, due to the peculiarity and vulnerability of these soils with a much reduced depth. The period of observation was carried out from autumn 2009 to autumn 2010. The sediment concentration of soil erosion under vines was recorded as 34.52 g/l while under pasture it was 4.66 g/l. In addition, the soil under vines showed the least amount of organic matter, which was 12 times lower than all other soil environments. The carbon dioxide (CO2) and total glomalin (TG) ratio to soil organic carbon (SOC) in this soil was 0.11 and 0.31 respectively. However, the soil under pasture contained a higher amount of organic matter and showed that the CO2 and TG ratio to SOC was 0.02 and 0.11 respectively indicating that the soil under pasture better preserves the soil carbon pool. A similar trend was found in the intermediate soils in the sequence of land use change and abandonment. Soil structural stability increased in the two soil fractions investigated (0.25-2.00 mm, 2.0-5.6 mm) especially in those soils that did not undergo periodical perturbations like wildfires. Soil quality indexes were obtained by using relevant physical and chemical soil parameters. Factor analysis carried out to study the relationship between all soil parameters allowed to related variables and environments and identify those areas that better contribute to soil quality towards others that may need more attention to avoid further degradation processes

Subjective perceptions as prognostic factors of time to fitness for work during a 4-year period after inpatient rehabilitation for orthopaedic trauma.

INTRODUCTION: Time to fitness for work (TFW) was measured as the number of days that were paid as compensation for work disability during the 4 years after discharge from the rehabilitation clinic in a population of patients hospitalised for rehabilitation after orthopaedic trauma. The aim of this study was to test whether some psychological variables can be used as potential early prognostic factors of TFW. MATERIAL AND METHODS: A Cox proportional hazards model was used to estimate the associations between predictive variables and TFW. Predictors were global health, pain at hospitalisation and pain decrease during the stay (all continuous and standardised by subtracting the mean and dividing by two standard deviations), perceived severity of the trauma and expectation of a positive evolution (both binary variables). RESULTS: Full data were available for 807 inpatients (660 men, 147 women). TFW was positively associated with better perceived health (hazard ratio [HR] 1.16, 95% confidence interval [CI] 1.13-1.19), pain decrease (HR 1.46, 95% CI 1.30-1.64) and expectation of a positive evolution (HR 1.50, 95% CI 1.32-1.70) and negatively associated with pain at hospitalisation (HR 0.67, 95% CI 0.59-0.76) and high perceived severity (HR 0.72, 95% CI 0.61-0.85). DISCUSSION: The present results provide some evidence that work disability during a four-year period after rehabilitation may be predicted by prerehabilitation perceptions of general health, pain, injury severity, as well as positive expectation of evolution.

Postnatal retention in HIV care : insight from the Swiss HIV Cohort Study over a 15-year observational period

OBJECTIVES: The aim of this study was to quantify loss to follow-up (LTFU) in HIV care after delivery and to identify risk factors for LTFU, and implications for HIV disease progression and subsequent pregnancies. METHODS: We used data on pregnancies within the Swiss HIV Cohort Study from 1996 to 2011. A delayed clinical visit was defined as > 180 days and LTFU as no visit for > 365 days after delivery. Logistic regression analysis was used to identify risk factors for LTFU. RESULTS: A total of 695 pregnancies in 580 women were included in the study, of which 115 (17%) were subsequent pregnancies. Median maternal age was 32 years (IQR 28-36 years) and 104 (15%) women reported any history of injecting drug use (IDU). Overall, 233 of 695 (34%) women had a delayed visit in the year after delivery and 84 (12%) women were lost to follow-up. Being lost to follow-up was significantly associated with a history of IDU [adjusted odds ratio (aOR) 2.79; 95% confidence interval (CI) 1.32-5.88; P = 0.007] and not achieving an undetectable HIV viral load (VL) at delivery (aOR 2.42; 95% CI 1.21-4.85; P = 0.017) after adjusting for maternal age, ethnicity and being on antiretroviral therapy (ART) at conception. Forty-three of 84 (55%) women returned to care after LTFU. Half of them (20 of 41) with available CD4 had a CD4 count < 350 cells/μL and 15% (six of 41) a CD4 count < 200 cells/μL at their return. CONCLUSIONS: A history of IDU and detectable HIV VL at delivery were associated with LTFU. Effective strategies are warranted to retain women in care beyond pregnancy and to avoid CD4 cell count decline. ART continuation should be advised especially if a subsequent pregnancy is planned.

The natural history of type II odontoid fractures in the elderly population. A retrospective study over a 14 years period.

Odontoid fractures are the most common cervical fractures in the adult population. They represent 9 to 18 % of all cervical fractures and the type II is the most common. The incidence of neurologic deficits (ND) in odontoid fractures varies between 3 to 25%. A recent study showed that patients with ND had a mortality rate increased by 4.72 times and a complication rate higher of 1.18 times. The most common complication in patients with ND was respiratory distress8. Surprisingly, although type II odontoid fractures are frequent cervical fractures, their natural history has been poorly described. Surgery for odontoid fractures is well described. However, there are so far guidelines based on class II and class III evidence only regarding indications for surgery and regarding surgical techniques. The class II guidelines recommend to consider surgical stabilization and fusion for type II odontoid in patients over 50 years of age. The class III recommendations are to first manage non-displaced odontoid type II fracture with external immobilization and that translation of 5mm or more is associated with a high rate of non- union with the conservative treatment and should be treated surgically.

Utilização do resíduo líquido de indústria de processamento de suco de laranja como meio de cultura de Penicillium citrinum: depuração biológica do resíduo e produção de enzima

Penicillium citrinum grown in orange juice processing wastes medium under continuous agitation was studied in order to establish optimal conditions (incubation period, incubation temperature, initial pH and nitrogen addition) for biomass and ribonuclease production, as well as, biological depuration of the wastes. Nitrogen addition to wastes medium increased the biomass and ribonuclease production and provides COD reduction. The soy meal shows to be the best nitrogen source. The conditions for a more favorable enzyme and biomass production and COD reduction were initial pH 6.0 and temperature of 27°C. The maximum value obtained for these parameters on optimal conditions of cultivation was 11 U/mL of enzyme, 4 mg/mL of biomass (dry matter) and 55% of COD reduction, in 96 hours of incubation.

Sexual foraging segregation in South American sea lions increases during the pre-breeding period in the La Plata River plume

Stable carbon and nitrogen isotopes in skin and bone of South American sea lions from Brazil and Uruguay were analysed to test the hypothesis that trophic overlap between the sexes is lower during the pre-breeding season than throughout the rest of the year. The isotopic values of skin and bone were used to infer the trophic relationships between the sexes during the pre-breeding period and year round, respectively. Prey species were also analysed to establish a baseline necessary for interpreting the stable isotope ratios of skin and bone. Standard ellipse areas, estimated using Bayesian inference in the SIBER routine of the SIAR package in R, suggested that males and females used a wide diversity of foraging strategies throughout the year and that no differences existed between the sexes. However, the diversity of foraging strategies was largely reduced during the pre-breeding period, with all the individuals of each sex adopting similar strategies, but with the two sexes differing considerably in stable isotope values and the ellipse areas of males and females not overlapping at all. Nevertheless, the results revealed a general increase in the consumption of pelagic prey by both sexes during the pre-breeding period. The progressive crowding of individuals in the areas surrounding the breeding rookeries during the pre-breeding period could lead to an increase in the local population density, which could explain the above reported changes.

Optimal latent period in a bacteriophage population model structured by infection-age

We study the lysis timing of a bacteriophage population by means of a continuously infection-age-structured population dynamics model. The features of the model are the infection process of bacteria, the death process, and the lysis process which means the replication of bacteriophage viruses inside bacteria and the destruction of them. The time till lysis (or latent period) is assumed to have an arbitrary distribution. We have carried out an optimization procedure, and we have found that the latent period corresponding to maximal fitness (i.e. maximal growth rate of the bacteriophage population) is of fixed length. We also study the dependence of the optimal latent period on the amount of susceptible bacteria and the number of virions released by a single infection. Finally, the evolutionarily stable strategy of the latent period is also determined as a fixed period taking into account that super-infections are not considered

A rota dos estudos sobre a cochonilha em Portugal e no Brasil no século XIX: caminhos desencontrados

For imparting an intense and long-lasting red to fabrics, cochineal remained in high demand during the eighteenth and nineteenth centuries. This period witnessed, accordingly, several initiatives aimed at producing the precious dye: publication of specialized texts, cultivation of the cactus and the insect from which the dye is extracted, and, also, attempts to obtain the secrets of production through espionage. The present paper analyses certain aspects of the measures adopted by the Portuguese government towards Brazil in this field. The work shows how people sought to take part in the network of cochineal production (yet they were unsuccessful most of the time).

Alternative technique for biodiesel quality control using an optical fiber long-period grating sensor

We report the use of an optical fiber sensor to measure the soybean oil concentration in samples obtained from the mixture of pure biodiesel and commercial soybean oil. The operation of the device is based on the long-period grating sensitivity to the surrounding medium refractive index, which leads to measurable modifications in the grating transmission spectrum. The proposed analysis method results in errors in the oil concentration of 0.4% and 2.6% for pure biodiesel and commercial soybean oil, respectively. Techniques of total glycerol, dynamic viscosity, density, and hydrogen nuclear magnetic resonance spectroscopy were also employed to validate the proposed method.

Bioaccumulation of metals in aquatic insects of streams located in areas with sugar cane cultivation

Streams located in areas of sugar cane cultivation receive elevated concentrations of metal ions from soils of adjacent areas. The accumulation of metals in the sediments results in environmental problems and leads to bioaccumulation of metal ions by the aquatic organisms. In the present study, bioaccumulation of the metals ions Al, Cd, Cr, Cu, Fe, Mg, Mn and Zn in aquatic insects in streams impacted by the sugar cane was evaluated. The results pointed out that the insects were contaminated by the sediment and that the collector organisms as Chironomus species accumulated higher concentration of metals than the predator organisms.

Incubation period of Hemileia vastatrix in coffee plants in Brazil simulated under climate change

Risk analysis of climate change on plant diseases has great importance for agriculture since it allows the evaluation of management strategies to minimize future damages. This work aimed to simulate future scenarios of coffee rust (Hemileia vastatrix) epidemics by elaborating geographic distribution maps using a model that estimates the pathogen incubation period and the output from three General Circulation Models (CSIRO-Mk3.0, INM-CM3.0, and MIROC3.2.medres). The climatological normal from 1961-1990 was compared with that of the decades 2020s, 2050s and 2080s using scenarios A2 and B1 from the IPCC. Maps were prepared with a spatial resolution of 0.5 × 0.5 degrees of latitude and longitude for ten producing states in Brazil. The climate variables used were maximum and minimum monthly temperatures. The maps obtained in scenario A2 showed a tendency towards a reduction in the incubation period when future scenarios are compared with the climatological normal from 1961-1990. A reduction in the period was also observed in scenario B1, although smaller than that in scenario A2.

Imazapyr root exudation from eucalypt seedlings cultivated in nutritive solution

Imazapyr has been used in Brazilian eucalypt cultivation for the maintenance of clearings and coppicing control in areas undergoing stand reform. However, inquiries have been made as to the final fate of the molecule. Imazapyr root exudation in eucalypt plants was evaluated through a bioassay under greenhouse conditions, by applying different herbicide doses (0.000, 0.375, 0.750, 1.125, and 1.500 kg ha-1 a.i.) on Eucalyptus grandis seedlings derived from vegetative propagation, hydroponically cultivated in 2.500 ml vases. Forty-day-old seedlings of the same clone were used as bioindicators, transplanted to the vases two days after herbicide application. After a period of 13 days of coexistence, the sprayed plants were removed and discarded; ten days later, the visual symptoms of toxicity were evaluated and the total dry biomass (aerial part and roots) of the bioindicators were determined. The lowest herbicide dose (0.375 kg ha-1 a.i.) affected the total biomass and growth, being most evident in the aerial part, with larger I50 for root dry biomass. The E. grandis seedlings exuded imazapyr, and/or its metabolites, in concentrations capable of affecting the growth of plants of the same species.