Increase in the thermostability of Bacillus sp. strain TAR-1 xylanase using a site saturation mutagenesis library

Site saturation mutagenesis library is a recently developed technique, in which any one out of all amino acid residues in a target region is substituted into other 19 amino acid residues. In this study, we used this technique to increase the thermostability of a GH10 xylanase, XynR, from Bacillus sp. strain TAR-1. We hypothesized that the substrate binding region of XynR is flexible, and that the thermostability of XynR will increase if the flexibility of the substrate binding region is decreased without impairing the substrate binding ability. Site saturation mutagenesis libraries of amino acid residues Tyr43–Lys115 and Ala300–Asn325 of XynR were constructed. By screening 480 clones, S92E was selected as the most thermostable one, exhibiting the residual activity of 80% after heat treatment at 80°C for 15 min in the hydrolysis of Remazol Brilliant Blue-xylan. Our results suggest that this strategy is effective for stabilization of GH10 xylanase.

Abbreviations: DNS: 3,5-dinitrosalicylic acid; RBB-xylan: Remazol Brilliant Blue-xylan     

Growth and Cadmium Phytoextraction by Swiss Chard,Maize, Rice,Noccaea caerulescens,and Alyssum murale in pH Adjusted Biosolids Amended Soils

Past applications of biosolids to soils at some locations added higher Cd levels than presently permitted. Cadmium phytoextraction would alleviate current land use constraints. Unamended farm soil, and biosolids amended farm and mine soils were obtained from a Fulton Co., IL biosolids management facility. Soils contained 0.16, 22.8, 45.3 mg Cd kg^–1 and 43.1, 482, 812 mg Zn kg^–1 respectively with initial pH 6.0, 6.1, 6.4. In greenhouse studies, Swiss chard (Beta vulgaris var. cicla), a Cd-accumulator maize (inbred B37 Zea mays) and a southern France Cd-hyperaccumulator genotype of Noccaea caerulescens were tested for Cd accumulation and phytoextraction. Soil pH was adjusted from ～5.5–7.0. Additionally 100 rice (Oryza sativa) genotypes and the Ni-hyperaccumulator Alyssum murale were screened for potential phytoextraction use.

Chard suffered phytotoxicity at low pH and accumulated up to 90 mg Cd kg^–1 on the biosolids amended mine soil. The maize inbred accumulated up to 45 mg Cd kg^–1 with only mild phytotoxicity symptoms during early growth at pH > 6.0. N. caerulescens did not exhibit phytotoxicity symptoms at any pH, and accumulated up to 235 mg Cd kg^–1 in 3 months. Reharvested N. caerulescens accumulated up to 900 mg Cd kg^–1 after 10 months. Neither Alyssum nor 90% of rice genotypes survived acceptably.

Both N. caerulescens and B37 maize show promise for Cd phytoextraction in IL and require field evaluation; both plants could be utilized for nearly continuous Cd removal. Other maize inbreds may offer higher Cd phytoextraction at lower pH, and mono-cross hybrids higher shoot biomass yields. Further, maize grown only for biomass Cd maximum removal could be double-cropped.     

Broad environmental tolerance of native root-nodule bacteria of Biserrula pelecinus indicate potential for soil fertility restoration

Background: Biserrula pelecinus is an annual legume native to the Mediterranean basin, found in pastureland, alone or in association with other legumes (Leguminosae) and grasses (Poaceae). It has been used in revegetation programmes of mining wastes showing phytoremediation potentials and thus becoming potentially highly attractive for plant ecology and restoration management of natural ecosystems.

Aims: To characterise native root-nodule bacteria isolated from B. pelecinus from the Iberian Peninsula, and to select suitable N fixers for field-application and soil rehabilitation.

Methods: Strains were isolated and molecularly identified by 16S rRNA amplification and sequencing. Strains were phenotypically characterised in different abiotic conditions (acidity, salinity and heavy metals) and tested for their ability to fix atmospheric N₂. The most suitable N fixers were applied in greenhouse experiments with B. pelecinus under different fertilization levels to assess their tolerance to fertilized and polluted soils, commonly encountered in restoration projects.

Results: B. pelecinus root-nodule isolates tolerated pH from 4.5 to 9.5 grew in saline conditions (2.5% of NaCl), and tolerated 50 µM of Al³⁺ and Mn²⁺. Three isolates efficient in N₂ fixation, relative to the reference Mesorhizobium strain, were considered excellent candidates for the amelioration of nutrient poor sites.

Conclusions: These results provide valuable information for the potential use in soil restoration of B. pelecinus in a wide-range of conditions, exploiting the natural variability of its root-nodule bacteria.     

Anti-diabetic effect of S-adenosylmethionine and α-glycerophosphocholine in KK-Ay mice

Six-week-old male KK-A^y mice received drinking water with S-adenosylmethionine (SAM), α-glycerophosphocholine (GPC), or SAM+GPC for 10 weeks. The serum glucose of SAM+GPC at 15 weeks old, total cholesterol of GPC at 12 weeks old, and triglyceride of GPC at 15 weeks old and of SAM at 16 weeks old were reduced. SAM+GPC reduced serum leptin and food intake.

Abbreviations: SAM: S-adenosylmethionine; GPC: α-glycerophosphocholine     

Dissolved Inorganic Carbon Inventory and CO2Sequestration in Ureolytic Biocalcification Process with Bacillus Pasteurii

The generation and depletion of dissolved inorganic carbon (DIC) in a close-environment ureolytic biocalcification process by Bacillus pasteurii was evaluated. Three experimental sets, each containing 50 mM urea, were amended with either 50 mM Ca²⁺ before incubation (set–I) or 100 mM Ca²⁺ after 24-h incubation (set-II) or no Ca²⁺ addition (urea control).

Extent of ureolysis was maximum in urea control set (88%), followed by set–II (66.4%) and set–I (35.2%). Out of total DIC generated from microbial metabolism and ureolysis in set–I (277.6 mg/l) and set–II (464.9 mg/l), only about 54.1 mg/l and 180.1 mg/l was precipitated as CaCO₃, whereas 189.3 mg/l and 231.3 mg/l DIC escaped into headspace, respectively. Increased time separation between ureolysis and calcification steps in set–II and higher dosage of Ca²⁺ resulted in synergistic improvement in DIC capture. In a reusability test, the spent supernatant from set–II could precipitate additional amount CaCO₃from CO₂saturated water_,which was twice as much as that of the fresh media control.     

Effect of grazing removal on aboveground vegetation and soil seed bank composition in sub-alpine grasslands of northern Iran

Background: Knowledge about vegetation and soil seed bank composition and the processes that contribute to vegetation recovery after the removal of heavy grazing is lacking in sub-alpine ecosystems.

Aims: In order to assess the effects of large herbivores on above-ground vegetation (AGV) and soil seed bank (SSB) characteristics, intensively sheep-grazed areas were compared to adjacent areas where grazing had been removed 10 years previously in a sub-alpine grassland of northern Iran.

Methods: A total of 40 4-m² (2 m × 2 m) plots were established in each treatment, and soil samples were collected from each plot within a depth of 10 cm. Plant species composition was determined for each plot during the flowering stage of herbaceous species in June 2011. The seedling emergence method was used to estimate SSB composition.

Results: A total of 45 species (23 annuals and 22 perennials) emerged from the soil samples of the grazed area, while the number of species emerged from the soil samples of the previously grazing area was 76 (37 annuals and 39 perennials). The removal of grazing led to a significant increase in species richness and seed density in the SSB. Species turnover of AGV was higher, and that of the SSB was similar for grazed areas compared with areas that were formerly grazed. Detrended correspondence analysis ordination of AGV composition showed a clearly separate structure between grazed plots and plots where grazing has been removed, while the segregation was less clear for SSB composition.

Conclusions: We concluded that restoration of locally degraded sites cannot rely on the SSB when grazing is stopped simultaneously and unvegetated gaps are colonised by vegetative growth rather than by seed.     

TRPA1 ion channel stimulation enhances cardiomyocyte contractile function via a CaMKII-dependent pathway

Rationale: Transient receptor potential channels of the ankyrin subtype-1 (TRPA1) are non-selective cation channels that show high permeability to calcium. Previous studies from our laboratory have demonstrated that TRPA1 ion channels are expressed in adult mouse ventricular cardiomyocytes (CMs) and are localized at the z-disk, costamere and intercalated disk. The functional significance of TRPA1 ion channels in the modulation of CM contractile function have not been explored.

Objective: To identify the extent to which TRPA1 ion channels are involved in modulating CM contractile function and elucidate the cellular mechanism of action.

Methods and Results: Freshly isolated CMs were obtained from murine heart and loaded with Fura-2 AM. Simultaneous measurement of intracellular free Ca²⁺ concentration ([Ca²⁺]_i) and contractility was performed in individual CMs paced at 0.3 Hz. Our findings demonstrate that TRPA1 stimulation with AITC results in a dose-dependent increase in peak [Ca²⁺]_i and a concomitant increase in CM fractional shortening. Further analysis revealed a dose-dependent acceleration in time to peak [Ca²⁺]_i and velocity of shortening as well as an acceleration in [Ca²⁺]_i decay and velocity of relengthening. These effects of TRPA1 stimulation were not observed in CMs pre-treated with the TRPA1 antagonist, HC-030031 (10 µmol/L) nor in CMs obtained from TRPA1^?/? mice. Moreover, we observed no significant increase in cAMP levels or PKA activity in response to TRPA1 stimulation and the PKA inhibitor peptide (PKI 14–22; 100 nmol/L) failed to have any effect on the TRPA1-mediated increase in CM contractile function. However, TRPA1 stimulation resulted in a rapid phosphorylation of Ca²⁺/calmodulin-dependent kinase II (CaMKII) (1–5 min) that correlated with increases in CM [Ca²⁺]_i and contractile function. Finally, all aspects of TRPA1-dependent increases in CM [Ca²⁺]_i, contractile function and CaMKII phosphorylation were virtually abolished by the CaMKII inhibitors, KN-93 (10 µmol/L) and autocamtide-2-related peptide (AIP; 20 µmol/L).

Conclusions: These novel findings demonstrate that stimulation of TRPA1 ion channels in CMs results in activation of a CaMKII-dependent signaling pathway resulting in modulation of intracellular Ca²⁺ availability and handling leading to increases in CM contractile function. Cardiac TRPA1 ion channels may represent a novel therapeutic target for increasing the inotropic and lusitropic state of the heart.     

Climate and fragmentation affect forest structure at the southern border of Amazonia

Background: The remaining forests in the extensive contact zone between southern Amazonia (seasonal rain forest) and the Cerrado (savanna) biomes are at risk due to intense land-use and climate change.

Aims: To explore the vulnerability of these transitional forests to changes in land use and climate, we evaluated the effects of fragmentation and climatic variables on forest structure.

Methods: We measured the diameter and height of 14,185 trees with diameter ≥10 cm at 24 forest plots distributed over an area of 25,000 km². For each plot, we obtained data on contemporary fragmentation and climatic variables.

Results: Forest structure variables (height, diameter, height:diameter allometry, biomass) varied significantly both within and among plots. The height, H:D and biomass of trees were positively correlated with annual precipitation and fragment area.

Conclusions: The association between forest structure and precipitation indicates that these forests plots are likely to be vulnerable to dry season intensification anticipated for the southern edge of the Amazon. Additionally, the reduction in the fragment area may contribute to reductions in forest biomass and tree height, and consequently ecosystem carbon stocks. Given the likely susceptibility of these forests, urgent conservation action is needed to prevent further habitat degradation.     

Fragmentation and loss of habitat: consequences for the fern communities in Atlantic forest remnants in Alagoas,north-eastern Brazil

Background: The reduction of forest areas and the fragmentation of remaining forest is a major threat to biodiversity and is one of the primary causes of species extinction in the Atlantic forest biome.

Aims: To evaluate the effects of the fragmentation and loss of habitat (size, edge effect, isolation and type of matrix) the richness, abundance, diversity and species composition of ferns in Atlantic forest fragments.

Methods: For each of 11 fragments, plots of 10 × 20 m² were compared in the interior and at the forest edge. Data were analysed by general linear models, or by the use of similarity indices for floristic composition.

Results: Eighty-eight species were recorded. Fragment size and type of environment (interior vs. edge) were related to the biological variables studied: large fragments and forest interior environments had larger populations of ferns, with a higher diversity. Floristic compositions were distinct for both the interior vs. edge environments and large vs. small fragments.

Conclusions: Although larger fragments present a higher number of species, the preservation of smaller fragments, which have a different suite of species from those found in the large fragments, is important to guarantee the maintenance of the highest number of fern species in the Atlantic forest. Small fragments are not merely subsets of the species occurring in the larger fragments. The turnover in the species composition between fragments of different size suggests high spatial variability in the study area.     

Relationships between chronotype,social jetlag,sleep, obesity and blood pressure in healthy young adults

Sleep disturbances, chronotype and social jetlag (SJL) have been associated with increased risks for major chronic diseases that take decades to develop, such as obesity, metabolic syndrome and cardiovascular disease. Potential relationships between poor sleep, chronotype and SJL as they relate to metabolic risk factors for chronic disease have not been extensively investigated. This prospective study examined chronotype, SJL and poor sleep in relation to both obesity and elevated blood pressure among healthy young adults.

SJL and objective sleep measures (total sleep time, sleep onset latency, wake after sleep onset and sleep efficiency) were derived from personal rest/activity monitoring (armband actigraphy) among 390 healthy adults 21–35 years old. Participants wore the device for 6–10 days at 6-month intervals over a 2-year period (n = 1431 repeated observations). Chronotypes were categorized into morning, intermediate and evening groups using repeated measures latent class analysis. Means of SJL and sleep measures among latent chronotype groups were compared using partial F-tests in generalized linear mixed models. Generalized linear mixed models also were used to generate odds ratios (ORs) with 95% confidence intervals (CIs) examining the relationship between repeated measures of chronotype, SJL, sleep and concurrent anthropometric outcome measures (body mass index, percentage of body fat, waist-to-hip ratio, waist-to-height ratio), systolic blood pressure and diastolic blood pressure.

Sleep latency ≥12 min was associated with increased odds of a high waist-to-height ratio (OR = 1.37; CI: 1.03–1.84). Neither chronotype nor SJL was independently associated with anthropometric outcomes or with blood pressure. Relationships between poor sleep and anthropometric outcomes or blood pressure varied by chronotype. Morning types with total sleep time <6 h, sleep efficiency <85% or wake after sleep onset ≥60 min were more likely to have an increased percentage of body fat, waist-to-hip ratio and waist-to-height ratio relative to those with an intermediate chronotype. Similarly, sleep latency ≥12 min was associated with increased odds of elevated systolic blood pressure (OR = 1.90; CI: 1.15–3.16, p_interaction = 0.02) among morning versus intermediate chronotypes. No relationships between poor sleep and obesity or elevated blood pressure were observed among evening chronotypes.

The results from this study among healthy young adults suggest that poor sleep among morning types may be more strongly associated with obesity and elevated blood pressure relative to those with an intermediate (neutral) chronotype. Sleep-related metabolic alterations among different chronotypes warrant further investigation.     

Lactobacillus gasseri PA-3 utilizes the purines GMP and guanosine and decreases their absorption in rats

Excessive intake of purine-rich foods elevates serum uric acid levels, making it a risk factor for hyperuricemia.

We hypothesized that lactic acid bacteria ingested with food might utilize purines and contribute to their decreased absorption in the intestines, thereby preventing hyperuricemia. We previously reported that Lactobacillus gasseri PA-3 (PA-3) incorporates adenosine/inosine and related purines and that oral ingestion of PA-3 reduced the absorption of these purines in rats. However, it is unclear whether PA-3 also decreases the absorption of other purines, such as guanosine 5′-monophosphate (GMP) and guanosine. This study investigated whether PA-3 incorporates GMP and guanosine and reduces their absorption in rats.

PA-3 incorporated both purines, with ¹⁴C-GMP uptake being greater than that of ¹⁴C-guanosine. Radioactivity in rat blood was significantly lower 30, 45, and 60 minutes after administration of ¹⁴C-GMP plus PA-3 than after administration of ¹⁴C-GMP alone and was significantly lower 15 minutes after administration of ¹⁴C-guanosine plus PA-3 than after administration of ¹⁴C-guanosine alone.

PA-3 incorporates GMP and guanosine in vitro. Oral administration of PA-3 with GMP and guanosine reduces the intestinal absorption of these purines in vivo. These findings, together with those of previous studies, indicate that PA-3 reduces the absorption of major purines contained in foods. PA-3 may also attenuate the excessive absorption of dietary purines in humans, protecting these individuals against hyperuricemia.     

Application of Diffusive Gradient in Thin Film to Estimate Available Copper in Soil Solution

In Chile, there are several agricultural areas with soils containing high levels of copper of both anthropic and geochemical origin. The diffusive gradient in thin film (DGT) technique is a promising tool for the evaluation of the bioavailability of metals in situ in different environmental systems. The objective of this study was the preparation, validation, and application of DGT to soil solutions and soil containing copper over 1000 mg kg^?1.

The results show that a resin gel thickness of 0.3 mm gives the best reproducibility and response in the absorption of copper by DGT. The amount of copper extracted in a period of 4 h by the devices from the soil solution corresponds to 13% of the total metal present in the solution.

The DGT allowed a more representative estimation of the amount of Cu available in the soil, more in agreement with the absence of symptoms of phytotoxicity in cultivated species. This shows that the determination of available Cu by DTPA must be handled cautiously because in soils with high Cu content the amount of metal that can have direct influence on absorption by the plant is overestimated.     

Observer error in sampling a rare plant population

Background: Estimation of abundance in vegetation sampling involving observers is almost always characterised by observer error, although such error is rarely reported.

Aims: To quantify observer error in population estimation of the rare plant species Physaria filiformis in Missouri, USA.

Methods: The abundance of P. filiformis was estimated within 25-m² plots by six trained observers with varying experience levels over 10 years. Observers assigned plots to six predefined density classes. A total of 477 plots were estimated annually, and actual counts were conducted on ca. 10% of the plots to assess per cent agreement of estimates with counts.

Results: Over a third of the estimates of plant abundance evaluated for accuracy (36.4%) deviated from exhaustive counts. The majority of the misestimates were underestimates by one density class (29.4%). The number and type of misestimates varied systematically with density class.

Conclusions: Observer error could be explained to some degree by variation in population density, but not by experience. It appears that inherent differences exist among observers that cannot be entirely compensated for by experience or training. Observer error in this system represents a systematic bias, and can be compensated for by use of correction factors, which would ideally be both density class-dependent and observer-specific.     

Sex differences in release of cardiac troponin T after endurance exercise

Context: Post-exercise cardiac troponin release has been extensively described in athletic groups but little attention has been given to any role of sex in mediating this phenomenon.

Objective: We compared the release of cardiac troponin T (cTnT) after endurance running in training-experience, biological-age and maturity-matched young male and female runners.

Materials and methods: Nineteen male (training history: 2.3?±?1.0?years; mean age: 16.1?±?1.2?years; Tanner stage: 3.7?±?0.6) and 19 female (training history: 2.2?±?1.0?years; mean age: 15.9?±?1.4?years; Tanner stage: 4.0?±?0.4) runners performed a 21?km run with “all-out” effort. Serum cTnT levels were assessed at pre-exercise (Pre-ex) and at 4?h post-exercise (Post-ex).

Results: At Pre-ex, cTnT concentrations were below the 99th percentile value (10?ng.l^?1) in 32/38 runners. Post-ex cTnT increased in all subjects but the response was substantially higher (p?<?0.05) in males [median (range): 210 (20–1360) ng.l^?1] than females [median (range): 80 (10–550) ng.l^?1]. At Post-ex, 95% (95% confidence interval: 75–99%) of males and 63% (95% confidence interval: 41–81%) of females (p?<?0.05) had cTnT concentrations above the cut-off for acute myocardial infarction.

Conclusions: The present data suggest that post-exercise cTnT elevation occurs in all runners but is augmented in young male compared to female athletes.     

Canopy cover and understory composition determine abundance of Vaccinium myrtillus L., a key plant for capercaillie (Tetrao urogallus), in subalpine forests in the Pyrenees

Background: The dwarf shrub Vaccinium myrtillus – with high cover, height, and fruit production – benefits capercaillie (Tetrao urogallus).

Aims: Our aim was to quantify landscape (e.g. elevation, geographic location, and precipitation), site (e.g. overstorey cover and stoniness) and very fine scale factors (e.g. spatial associations in the understorey) that affect cover, height, and fruit production of V. myrtillus in subalpine forests in thePyrenees, with understorey usually dominated by Rhododendron ferrugineum.

Methods: We sampled 155 plots (0.5 m × 5 m) in six sites. For each plot, in the understorey layer, we assessed species cover, height for R. ferrugineum and V. myrtillus, number of total fruits in V. myrtillus, and spatial associations among V. myrtillus and the remaining cover types.

Results: Overstorey cover negatively influenced V. myrtillus cover, its height, and particularly, the number of fruits, which was also negatively influenced by R. ferrugineum cover. Associations between R. ferrugineum and V. myrtillus were site dependent, while V. myrtillus showed mostly positive associations with grasses and mosses.

Conclusions: Reducing overstorey and R. ferrugineum cover has the strongest positive effect on increasing V. myrtillus fruit production, but with additional positive effects on V. myrtillus cover and height. Increases in grass and moss coverage could favour V. myrtillus.     

Thrombin generation is increased in patients with Clostridium difficile colitis – a pilot study

Background: There are only limited data in the literature on the thrombotic risk of patients with Clostridium difficile (CD) colitis, although this disease is widespread throughout the world.

Objective: The aim of this study was to explore thrombin generation in these patients – the best way to evaluate their coagulation.

Methods: A prospective observational study was conducted during 15 months on hospitalized patients with CD colitis. Thrombin generation was performed in platelet-poor plasma using a Ceveron^® alpha analyzer and was compared with a group of volunteer control subjects.

Results: Thirty-three patients and 51 control subjects were enrolled in the study. Two biomarkers – mean velocity index and peak thrombin – were significantly higher in patient group, compared to the control subjects (p?=?0.010, respectively, p?=?0.0395). This pattern of thrombin generation suggests that patients with CD colitis without septic shock have a potential thrombotic risk. The mean velocity index significantly correlated with the estimated related risk of death according to the Charlson age-comorbidity index.

Conclusions: The higher values of thrombin generation suggest that CD colitis increases the thromboembolic risk. The pattern of thrombin generation could identify patients with particularly higher thromboembolic risk. They are potential candidates for thromboprophylaxis strategies and monitorization.     

Circadian gene methylation in rotating-shift nurses: a cross-sectional study

Investigating the methylation status of the circadian genes may contribute to a better understanding of the shift work-related circadian disruption in individuals exposed to artificial light at night. In the present study, we determined the methylation status of the circadian genes associated with a shift work pattern among nurses and midwives participating in a cross-sectional study in Lodz, Poland.

Quantitative methylation polymerase chain reaction assays were used to assess promoter CpG methylation in PER1, PER2, PER3, CRY1, CRY2, BMAL1, CLOCK, and NPAS2 in genomic DNA from whole blood of 347 women having a rotating-shift work schedule and 363 women working days only. The percentage of methylated reference (PMR) was assessed using fluorescent probes for PER1, PER2, PER3, CRY1, and NPAS2, and the percentage of gene methylation, as the methylation index (MI), using two sets of primers for BMAL1, CLOCK, and CRY2.

We tested the possible association between current and lifetime rotating night-shift work characteristics and circadian gene methylation by using proportional odds regression model with blood DNA methylation, categorized into tertiles, and adjusted for age, current smoking status, folate intake and blood collection time. The findings indicated that CpG methylation in PER2 promoter was significantly decreased (P < 0.004) among nurses and midwives currently working rotating shifts, as compared with day-working nurses and midwives. The lower percentage of PER2 methylation was associated with a higher monthly frequency of current night duties (2–7 night shifts, and eight or more night shifts per month) (P = 0.012) and was associated at borderline significance (P = 0.092) with the lifetime duration of shift work (>10 ≤ 20 years and >20 ≤ 43 years of rotating-shift work) among nurses and midwives (N = 710). Moreover, women with a longer lifetime duration of shift work presented a lower status of PER1 methylation (P = 0.040) than did the women with up to 10 years of rotating-shift work. Long lifetime duration of shift work (> 10 years) among current rotating night-shift workers (N = 347) was associated with BMAL1 hypomethylation (P = 0.013).

Among eight of the investigated circadian genes, only PER1, PER2, and BMAL1 showed differential methylation attributable to the rotating-shift work of nurses and midwives. The findings on blood-based DNA methylation in the circadian genes may provide a better insight into the mechanistic principles underlying the possible health effects of night-shift work but these should be verified in further studies recruiting larger populations of shift workers.     

From evergreen to deciduous tropical forests: how energy–water balance,temperature, and space influence the tree species composition in a high diversity region

Background: Understanding floristic and geographic patterns in one of the most biodiverse regions in the world – the Atlantic forest of eastern Bahia, Brazil – can identify the drivers of diversity in tropical forests and provide useful information for biological conservation.

Aims: To understand the role of both climate and geographical location on variation in tree species composition in a region characterised by an abrupt transition from wet forests to semi-arid thorn-woodlands. To test whether a regional classification of forests according to elevation belts and leaf flush pattern is consistent with floristic composition.

Methods: We submitted 14,094 tree species occurrence records and 31 geo-climatic variables prepared for 57 sites in eastern Bahia, Brazil, to multivariate and regression analyses and variance partitioning.

Results: Climate and space were both significantly (P ≤ 0.05) contributing to explaining floristic variations. Actual evapotranspiration, duration of water deficit, and minimum temperature of coldest month were the main predictors. Floristic differences were significant except when comparing evergreen lower plains and upper plains forests.

Conclusions: Although distance among sites may play an important role, species composition is chiefly influenced by environmental gradients. This highlights environmental heterogeneity as a key factor in the planning of biodiversity conservation in tropical forests.     

The temporal pattern of immune and inflammatory proteins prior to a recurrent coronary event in post-acute coronary syndrome patients

Purpose: We assessed the temporal pattern of 29 immune and inflammatory proteins in post-acute coronary syndrome (ACS) patients, prior to the development of recurrent ACS.

Methods: High-frequency blood sampling was performed in 844 patients admitted for ACS during one-year follow-up. We conducted a case-control study on the 45 patients who experienced reACS (cases) and two matched event-free patients (controls) per case. Olink Proteomics’ immunoassay was used to obtain serum levels of the 29 proteins, expressed in an arbitrary unit on the log2-scale (Normalized Protein eXpression, NPX). Linear mixed-effects models were applied to examine the temporal pattern of the proteins, and to illustrate differences between cases and controls.

Results: Mean age was 66?±?12 years and 80% were men. Cases and controls had similar baseline clinical characteristics. During the first 30 days, and after multiple testing correction, cases had significantly higher serum levels of CXCL1 (difference of 1.00 NPX, p?=?0.002), CD84 (difference of 0.64 NPX, p?=?0.002) and TNFRSF10A (difference of 0.41 NPX, p?<?0.001) than controls. After 30 days, serum levels of all 29 proteins were similar in cases and controls. In particular, no increase was observed prior to reACS.

Conclusions: Among 29 immune and inflammatory proteins, CXCL1, CD84 and TNFRSF10A were associated with early reACS after initial ACS-admission.     

Bright light exposure before bedtime impairs response inhibition the following morning: a non-randomized crossover study

Introduction: Bright light exposure in the late evening can affect cognitive function the following morning either by changing the biological clock and/or disturbing sleep, but the evidence for this effect is scarce, and the underlying mechanism remains unknown. In this study, we first aimed to evaluate the effect of bright light exposure before bedtime on frontal lobe activity the following morning using near-infrared spectroscopy (NIRS) during a Go/NoGo task. Second, we aimed to evaluate the effects of bright light exposure before bedtime on polysomnographic measures and on a frontal lobe function test the following morning.

Methods: Twenty healthy, young males (mean age, 25.5 years) were recruited between September 2013 and August 2014. They were first exposed to control light (150 lux) before bedtime (from 20:00 h to 24:00 h) for 2 days and then to bright light (1,000 lux) before bedtime for an additional 5 days. We performed polysomnography (PSG) on the final night of each light exposure period (on nights 2 and night 7) and performed NIRS, which measures the concentrations of oxygenated and deoxygenated hemoglobin (OxyHb and DeoxyHb, respectively), coupled with a Go/NoGo task the following morning (between 09:30 h and 11:30 h). The participants also completed frontal lobe function tests the following morning.

Results: NIRS showed decreased hemodynamic activity (lower OxyHb and a tendency toward higher DeoxyHb concentration) in the right frontal lobe during the NoGo block after 1000-lux light exposure compared with that during the NoGo block after 150-lux light exposure. The commission error rate (ER) during the Go/NoGo task was higher after 1000-lux light exposure than that during the Go/NoGo task after 150-lux light exposure (1.24 ± 1.09 vs. 0.6 ± 0.69, P = 0.002), suggesting a reduced inhibitory response.

Conclusion: This study shows that exposure to bright light before bedtime for 5 days impairs right frontal lobe activation and response inhibition the following morning.