Epigenetic control on transcription of vernalization genes and whole-genome gene expression profile induced by vernalization in common wheat.

Vernalization is necessary for winter wheat to flower. However, it is unclear whether vernalization is also required for spring wheat, which is frequently sown in fall, and what molecular mechanisms underlie the vernalization response in wheat varieties. In this study, we examined the molecular mechanisms that regulate vernalization response in winter and spring wheat varieties. For this purpose, we determined how major vernalization genes (VRN1, VRN2, and VRN3) respond to vernalization in these varieties and whether modifications to histones play a role in changes in gene expression. We also identified genes that are differentially regulated in response to vernalization in winter and spring wheat varieties. We found that in winter wheat, but not in spring wheat, VRN1 expression decreases when returned to warm temperature following vernalization. This finding may be associated with differences between spring and winter wheat in the levels of tri-methylation of lysine 27 on histone H3 (H3K27me3) and tri-methylation of lysine 4 on histone H3 (H3K4me3) at the VRN1 gene. Analysis of winter wheat transcriptomes before and after vernalization revealed that vernalization influences the expression of several genes, including those involved in leucine catabolism, cysteine biosynthesis, and flavonoid biosynthesis. These findings provide new candidates for further study on the mechanism of vernalization regulation in wheat.

Mechanisms of glioblastoma replication: Ca2+ flares and Cl- currents.

Glioblastoma (GBM) is amongst the deadliest types of cancers, with no resolutive cure currently available. GBM cell proliferation in the patient's brain is a complex phenomenon controlled by multiple mechanisms. The aim of this study was to determine whether the ionic fluxes controlling cell duplication could represent a target for GBM therapy. In this work, we combined multi-channel Ca2+ and Cl- imaging, optical tweezers, electrophysiology and immunohistochemistry to describe the role of ion fluxes in mediating the cell volume changes that accompany mitosis of U87 GBM cells. We identified three main steps: (i) in round GBM cells undergoing mitosis, during the transition from anaphase to telophase and cytokinesis, large Ca2+ flares occur, reaching values of 0.5-1 µM; (ii) these Ca2+ flares activate Ca2+-dependent Cl- channels, allowing the entry of Cl- ions; (iii) to maintain osmotic balance, GBM cells swell to complete mitosis. This sequence of steps was validated by electrophysiological experiments showing that Cl- channels are activated either directly or indirectly by Ca2+, and by additional live-cell imaging experiments. Cl- channel blockers with different molecular structures, such as niflumic acid and carbenoxolone, blocked GBM replication by arresting GBM cells in a round configuration. These results describe the central role of Ca2+ flares and Cl- fluxes during mitosis and show that inhibition of Ca2+-activated Cl- channels blocks GBM replication, opening the way to new approaches for the clinical treatment of GBM. Implications: Our work identifies ionic fluxes occurring during cell division as targets for devising novel therapies for the glioblastoma treatment.

Colloidal gold and fluorescent immunochromatographic test strips for canine parvovirus detection.

Canine parvovirus (CPV) is an acute and highly infectious virus causing disease in puppies and, thus, affecting the global dog industry. The current CPV detection methods are limited by their sensitivity and specificity. Hence, the current study sought to develop a rapid, sensitive, simple, and accurate immunochromatographic (ICS) test to detect and control the spread and prevalence of CPV infection. More specifically, 6A8, a monoclonal antibody (mAb) with high specificity and sensitivity, was obtained by preliminary screening. The 6A8 antibody was labelled with colloidal gold particles. Subsequently, 6A8 and goat anti-mouse antibodies were coated onto a nitrocellulose membrane (NC) as the test and control lines, respectively. Furthermore, 6A8 and rabbit IgG antibodies were labelled with fluorescent microspheres and evenly sprayed onto a glass fibre membrane. Both strips could be prepared in 15 min with no noticeable cross-reactivity with other common canine intestinal pathogens. The strips were simultaneously used to detect CPV in 60 clinical samples using real-time quantitative PCR, hemagglutination, and hemagglutination inhibition assays. The colloidal gold (fluorescent) ICS test strip was stable for 6 (7) and 4 (5) months at 4 °C and room temperature (18-25 °C). Both test strips were easy to prepare and rapidly detected CPV with high sensitivity and specificity. Moreover, the results were easily interpretable. This study establishes a simple method for two CPV diseases, colloidal gold and fluorescent immunochromatographic (ICS) test strips. KEY POINTS: • CPV test strips do not exhibit cross-reactivity with other canine intestinal pathogens. • The strips are stable for months at 4 °C and at room temperature (18-25 °C). • These strips are a promising approach for the timely diagnosis and treatment of CPV.

Differential regulation of phosphorylation, structure, and stability of circadian clock protein FRQ isoforms.

Neurospora crassa is an important model organism for circadian clock research. The Neurospora core circadian component FRQ protein has two isoforms, large FRQ (l-FRQ) and small FRQ (s-FRQ), of which l-FRQ bears an additional N-terminal 99-amino acid fragment. However, how the FRQ isoforms operate differentially in regulating the circadian clock remains elusive. Here, we show l-FRQ and s-FRQ play different roles in regulating the circadian negative feedback loop. Compared to s-FRQ, l-FRQ is less stable and undergoes hypophosphorylation and faster degradation. The phosphorylation of the C-terminal l-FRQ 794-aa fragment was markedly higher than that of s-FRQ, suggesting the l-FRQ N-terminal 99-aa region may regulate the phosphorylation of the entire FRQ protein. Quantitative label-free LC/MS analysis identified several peptides that were differentially phosphorylated between l-FRQ and s-FRQ, which were distributed in FRQ in an interlaced fashion. Furthermore, we identified two novel phosphorylation sites, S765 and T781; mutations S765A and T781A showed no significant effects on conidiation rhythmicity, although T781 conferred FRQ stability. These findings demonstrate that FRQ isoforms play differential roles in the circadian negative feedback loop and undergo different regulations of phosphorylation, structure, and stability. The l-FRQ N-terminal 99-aa region plays an important role in regulating the phosphorylation, stability, conformation, and function of the FRQ protein. As the FRQ circadian clock counterparts in other species also have isoforms or paralogues, these findings will also further our understanding of the underlying regulatory mechanisms of the circadian clock in other organisms based on the high conservation of circadian clocks in eukaryotes.

Observation and identification of autofluorescent urine crystals may be linked to a sign of urolithiasis.

Urolithiasis is a common disease of the urinary system. Its recurrence rate is high and may increase medical expenses. Urine stones are composed of urine crystals and other impurities. We discovered the existence of autofluorescence in some of the urine crystals, especially in urolithiasis patients. The fluorescent molecule existed in urine crystals was verified and identified. We have applied micro-Raman and fluorescence microscopy to classify the urine crystals, used confocal laser scanning microscopy (CLSM) to examine the 3D images and spectra of autofluorescence in crystals, used Fourier-transform infrared spectroscopy (FTIR) and mass spectrometry (MS) to identify the type of fluorophore in the autofluorescent urine crystals in urine. Riboflavin was identified as one of the major fluorophores in these autofluorescent urine crystals. The prevalence rates of the autofluorescent crystals in urolithiasis patients and subjects without the history of urolithiasis were to gather statistics. We observed that 80% of urolithiasis patients had autofluorescent crystals. Contrastingly, such crystals existed in only 7% of subjects without the history of urolithiasis. The presence of autofluorescent urine crystals may be linked to a sign of urolithiasis.

Comprehensive detrimental effects of a simulated frequently shifting schedule on diurnal rhythms and vigilance.

Accumulating data have demonstrated that shift work causes a disturbance in circadian rhythms, which is detrimental to physiology and performance. However, the detailed effects of shift work and especially the underlying mechanisms remain to be further investigated. Frequently shifting schedules are widely used in industries, e.g., maritime tasks, oil mining, and aviation. In this work, we investigated the physiological changes and vigilance of 12 subjects who lived on a 30-day frequent shift working schedule in a confined environment, which mimics the common maritime schedules. Elevated and decreased cortisol levels were observed at different stages during the shift, suggesting the occurrence of stress and fatigue. The results of the Karolinska Sleepiness Scale (KSS) indicate increased sleepiness and a changed pattern of the rhythmicity of sleepiness during the shift. The tests of the Psychomotor Vigilance Task (PVT) reveal that the shift led to a continuously decreasing alertness as the shift working schedule progressed, which is prevalently due to the increasingly slower reaction speed. The PVT time-out errors were significantly increased in the early period but decreased in the late period. In addition, we found recoupling of the correlations between multiple physiological and cognitive variables. For instance, heartbeat rate (HR) and breath rate (BR) showed moderate correlations in the control and early periods but little in the late period. Together, these results reveal substantial alterations in diurnal rhythms, affected vigilance and changed coupling of the correlations of diurnal rhythms, physiology and cognition caused by a shift schedule. Our findings may help in the recognition of the detrimental effects of such working schedules and provide clues for the development of potential mitigations.

Mechanotransduction in hippocampal neurons operates under localized low picoNewton forces.

There is growing evidence suggesting that mechanical properties of CNS neurons may play an important regulatory role in cellular processes. Here, we employ an oscillatory optical tweezers (OOT) to exert a local indentation with forces in the range of 5-50 pN. We found that single local indentation above a threshold of 13 ± 1 pN evokes a transient intracellular calcium change, whereas repeated mechanical stimulations induce a more sustained and variable calcium response. Importantly, neurons were able to differentiate the magnitude of mechanical stimuli. Chemical perturbation and whole-cell patch clamp recordings suggest that mechanically evoked response requires the influx of extracellular calcium through transmembrane ion channels. Moreover, we observed a mechanically evoked activation of the CAMKII and small G protein RhoA. These results all together suggest that mechanical signaling among developed neurons fully operates in neuronal networks under physiological conditions.

Identification and characterization of An-4, a potential quantitative trait locus for awn development in rice.

BACKGROUND: Awn of rice is an important domestication trait closely associated with yield traits. Therefore, the identification of genes for awn development is of great significance for the elucidation of molecular mechanism of awn development and the genetic improvement of yield traits in rice. RESULTS: In this study, using chromosome segment substitution lines (CSSLs) derived from a long-awned Guangxi common wild rice (GXCWR, Oryza rufipogon Griff.) and a short-awned indica cultivar 9311, we identified An-4, a potential quantitative trait locus (QTL) for awn development. Then, An-4 was fine mapped into a 56-kb region of chromosome 2, which contained four annotated genes. Among these four annotated genes, Os02g0594800 was concluded to be the potential candidate gene for An-4. An-4 exhibited pleiotropic effects on awn development and several yield traits. Scanning electron microscopy (SEM) analysis showed that An-4 significantly promoted awn development at Sp7 and Sp8 stage of spikelet development. Transcriptome analysis suggested that An-4 might influence the development of awn by regulating the expression of genes related to growth, developmental process, channel regulation and extracellular region. By contrast to those of 9311, the expression level of OsRR5 in CSSL128 was significantly down-regulated, whereas the expression levels of OsCKX2 and OsGA2ox5 in CSSL128 were significantly up-regulated. In addition, our study showed that An-4 had additive effects with other genes for awn development, such as An-1, An-2/LABA1 and An-3/GAD1/RAE2. CONCLUSIONS: The identification of An-4 lays a foundation for cloning of An-4 and further elucidation of the molecular mechanism of awn development. Moreover, the identification of favorable allelic variation of An-4 from 9311 will be useful to improve rice yield traits.

Study on the complexation of heavy metals onto biogas slurry DOM using two-dimensional correlation spectroscopy combined with the log-transformed synchronous fluorescence spectroscopy.

The fluorescent components of dissolved organic matter (DOM) in biogas slurry can react with heavy metals (HMs) and affect the migration, transformation, toxicity, and bioavailability of HMs in soil. Fluorescence quenching titration combined with two-dimensional correlation spectroscopy (2D-COS) can reveal the binding mechanism between HMs and different fluorescent components of biogas slurry DOM. The logarithmic-transformed (log-transformed) 2D-COS can be used to decrease the difference in the fluorescence intensity between low-intensity and high-intensity fluorophores that provides a better insight into the binding mechanism between biogas slurry DOM and HMs. Synchronous maps suggest that protein-like substances are more susceptive to the variation of the concentration of metal ions than fulvic-like substances. Asynchronous maps show that the preferential bonding of Cu(II) and Cr(III) to humic-like substances can be found in the biogas slurry DOM, as well as Fe(III) and Pb(II) to protein-like materials. DOM-Cu(II) may lead to an increasing risk of the migration of Cu(II) from soil to water environment due to the low log K values in the range from 2.93 to 3.46. Protein-like substances can also increase the environmental risk of HMs when these low-stable complexes occur migration and transformation. The potential environmental risk of protein-like with HMs follows the order: Pb(II) > Cu(II) > Cr(III). Here we demonstrate that the log-transformed 2D-COS can also identify fluorescence components at longer wavelength with relatively low content and reveals their preferential binding sequence and the number of binding sites. The study on the complexation between biogas slurry DOM and HMs provides a scientific basis for the environmental chemical behavior of HMs after the application of biogas slurry in agricultural soils.

The Resonance and Adaptation of Neurospora crassa Circadian and Conidiation Rhyth ms to Short Light-Dark Cycles.

Circadian clocks control the physiological and behavioral rhythms to adapt to the environment with a period of ~24 h. However, the influences and mechanisms of the extreme light/dark cycles on the circadian clock remain unclear. We showed that, in Neurospora crassa, both the growth and the microconidia production contribute to adaptation in LD12:12 (12 h light/12 h dark, periodically). Mathematical modeling and experiments demonstrate that in short LD cycles, the expression of the core clock protein FREQUENCY was entrained to the LD cycles when LD > 3:3 while it free ran when T ≤ LD3:3. The conidial rhythmicity can resonate with a series of different LD conditions. Moreover, we demonstrate that the existence of unknown blue light photoreceptor(s) and the circadian clock might promote the conidiation rhythms that resonate with the environment. The ubiquitin E3 ligase FWD-1 and the previously described CRY-dependent oscillator system were implicated in regulating conidiation under short LD conditions. These findings shed new light on the resonance of Neurospora circadian clock and conidiation rhythms to short LD cycles, which may benefit the understandings of both the basic regulatory aspects of circadian clock and the adaptation of physiological rhythms to the extreme conditions.

Effect of terminal temperature on the morphology and potentially toxic metals concentrations of biochars derived from paper and kitchen waste.

This study investigated the morphology and potentially toxic metal concentrations of paper waste-based biochar (PB) and kitchen waste-based biochar (KB) obtained at 500 and 700 °C. The morphology and potentially toxic metals (Cr, Mn, Cu, Cd, Pb, Zn, Ag, and Ba) concentrations in the biochars were determined by SEM and FT-IR analysis. The Cr, Mn, Cu, and Cd concentrations in PB were low, while the Ba content was relatively high at 0.1 mg∙kg-1. An increase in the terminal temperature led to an increase in the concentrations of Fe/Mn oxide-bound potentially toxic metals of PB, and a decrease in the concentrations of organic matter-bound potentially toxic metals. The Fe/Mn oxide-bound Cr, Mn, Cu, Pb, and Zn concentrations of KB decreased with an increase in the terminal temperature. Therefore, increasing the terminal temperature could reduce the bioavailability of potentially toxic metals in PB and KB. The environmental risk of the different biochars when used for soil remediation was assessed by the potential ecological risk index (RI), and a case study of a Tibetan soil was also conducted. The potentially toxic metal concentrations leached from both PB and KB were lower than the relevant standards. The findings showed that both PB and KB can be safely used for soil remediation.

Calcium flares and compartmentalization in rod photoreceptors.

Rod photoreceptors are composed of a soma and an inner segment (IS) connected to an outer segment (OS) by a thin cilium. OSs are composed of a stack of ∼800 lipid discs surrounded by the plasma membrane where phototransduction takes place. Intracellular calcium plays a major role in phototransduction and is more concentrated in the discs, where it can be incorporated and released. To study calcium dynamics in rods, we used the fluorescent calcium dye CaSiR-1 AM working in the near-infrared (NIR) (excitation at 650 and emission at 664 nm), an advantage over previously used dyes. In this way, we investigated calcium dynamics with an unprecedented accuracy and most importantly in semidark-adapted conditions. We observed light-induced drops in [Ca2+]i with kinetics similar to that of photoresponses recorded electrophysiologically. We show three properties of the rods. First, intracellular calcium and key proteins have concentrations that vary from the OS base to tip. At the OS base, [Ca2+]i is ∼80 nM and increases up to ∼200 nM at the OS tip. Second, there are spontaneous calcium flares in healthy and functional rod OSs; these flares are highly localized and are more pronounced at the OS tip. Third, a bright flash of light at 488 nm induces a drop in [Ca2+]i at the OS base but often a flare at the OS tip. Therefore, rod OSs are not homogenous structures but have a structural and functional gradient, which is a fundamental aspect of transduction in vertebrate photoreceptors.

Effect of irrigation water system's distribution on rice cadmium accumulation in large mild cadmium contaminated paddy field areas of Southwest China.

There are large areas of moderately Cd-contaminated rice paddies (Cd content was less than risk intervention value) in southwest China, under natural conditions, the effect of irrigation water system's distribution on the Cd contamination in soil and rice is still less accurate. In this study, a survey of paired soil-rice (n = 1520) samples was conducted in a large paddy of about 7000 ha in southwest China that originated from the same parent material and grew with the same rice varieties. Specially, three representative characteristic regions (area A, B, C) were selected from north to south to thoroughly investigate the reasons for pollution characteristics. Background soil, irrigation water and sediment, atmospheric deposition, fertilizer were sampled to study the causes of pollution. Results showed the biological accumulation factor (BAF = C (rice Cd)/C (soil Cd)) of area C in the south reached 1.34, which was about 8 times higher than that of area A in the north. The uneven distribution of irrigation water due to geographical reasons was the most important factor leading to this pollution characteristics. The Cd content in soil of north was much higher than that in the south due to the history of Cd-contaminated irrigation and background content of Cd. During farmland formation, river impinges resulted in a gradual decrease in both Cd content and pH in the background soil from north to south. Both of historical polluters and major irrigation systems were distributed in area A. However, when sewage irrigation stopped about 30 years ago, long-term weakly alkaline irrigation increased the pH of the soil from acidic to neutral in area A. Meanwhile, flooding irrigation in area A reduced the absorption of Cd by roots compared area C where was rarely flooded during the rice planting process due to the lack of irrigation water.

Mechanosensitivity is an essential component of phototransduction in vertebrate rods.

Photoreceptors are specialized cells devoted to the transduction of the incoming visual signals. Rods are able also to shed from their tip old disks and to synthesize at the base of the outer segment (OS) new disks. By combining electrophysiology, optical tweezers (OTs), and biochemistry, we investigate mechanosensitivity in the rods of Xenopus laevis, and we show that 1) mechanosensitive channels (MSCs), transient receptor potential canonical 1 (TRPC1), and Piezo1 are present in rod inner segments (ISs); 2) mechanical stimulation-of the order of 10 pN-applied briefly to either the OS or IS evokes calcium transients; 3) inhibition of MSCs decreases the duration of photoresponses to bright flashes; 4) bright flashes of light induce a rapid shortening of the OS; and 5) the genes encoding the TRPC family have an ancient association with the genes encoding families of protein involved in phototransduction. These results suggest that MSCs play an integral role in rods' phototransduction.

Impacts of a novel strain QY-1 allied with chromium immobilizing materials on chromium availability and soil biochemical properties.

In-situ passivation of soil chromium (Cr) contamination based on chemical and biological passivators has been widely concerned, however, the cooperative effect of two types of passivators on Cr passivation and soil properties was little investigated. In this study, nano zero valent iron (nZVI) and humic acid (HA) as the chemical passivators were selected and were combined with a novel Cr resistant strain QY-1 to study these two points. Results demonstrated that the combination was more effective in Cr immobilization, among which, HA + QY-1 had the highest passivation rate (82.83%), followed by nZVI + QY-1. HA + QY-1 alleviated soil Cr stress most efficiently as its soil relevant fertility indicators, microbial quantity, respiration and seed gemination rate significantly increased. On the contrary, nZVI decreased soil respiration and microbial abundance, but the addition of QY-1 could relieve this phenomenon. The results highlighted the ability of HA + QY-1 to remediate Cr contaminated soil and improve soil stability.

Sleep deprivation and a non-24-h working schedule lead to extensive alterations in physiology and behavior.

Most organisms on Earth possess circadian rhythms in their physiology and behaviors that allow them to resonate with the cycling environment over a 24-h period. However, in human society, a substantial quantity of jobs requires non-24-h working and rest or shift schedules, which causes more or less misalignment in circadian rhythms and disorders as a consequence. In this work, we conducted a sleep deprivation (SD) and non-24-h working and rest schedule (8 h on and 4 h off) experiment over 10 d in total and measured the changes in a series of physiologic and cognitive parameters. The results show that although the subjects could sleep during the schedule, their sleepiness increased significantly. Actigraphy data suggest that a 12-h schedule might result in chronic SD. Along with the increased sleepiness revealed by the Karolinska Sleepiness Scale questionnaire, the neurobehavioral psychomotor vigilance test data reveal that, compared with the control period, the reaction time of the subjects was significantly delayed. The saliva insulin levels were significantly changed in the morning in SD and non-24-h cycles. Salivary biochemical parameters were also altered, including aspartate aminotransferase and K+. 16S rRNA-based analysis of the salivary microbiota showed differentially changed patterns in bacteria composition and concentration. Together, these data demonstrate that an abnormal working and rest schedule might produce comprehensive interference with circadian rhythms, metabolism, and cognition.-Ma, H., Li, Y., Liang, H., Chen, S., Pan, S., Chang, L., Li, S., Zhang, Y., Liu, X., Xu, Y., Shao, Y., Yang, Y., Guo, J. Sleep deprivation and a non-24-h working schedule lead to extensive alterations in physiology and behavior.

The effect of two different biochars on remediation of Cd-contaminated soil and Cd uptake by Lolium perenne.

Biochar can be widely used to reduce the bioavailability of heavy metals in contaminated soil because of its adsorption capacity. But there are few studies about the effects of biochar on cadmium uptake by plants in soil contaminated with cadmium (Cd). Therefore, an incubation experiment was used to investigate the effects of rice straw biochar (RSBC) and coconut shell biochar (CSBC) on Cd immobilization in contaminated soil and, subsequently, Cd uptake by Lolium perenne. The results showed that the microbial counts and soil enzyme activities were significantly increased by biochar in Cd-contaminated soil, which were consistent with the decrease of the bioavailability of Cd by biochar. HOAc-extractable Cd in soil decreased by 11.3-22.6% in treatments with 5% RSBC and by 7.2-17.1% in treatments with 5% CSBC, respectively, compared to controls. The content of available Cd in biochar treatments was significantly lower than in controls, and these differences were more obvious in treatment groups with 5% biochar. The Cd concentration in L. perenne reduced by 4.47-26.13% with biochar. However, the biomass of L. perenne increased by 1.35-2.38 times after adding biochar amendments. So, Cd uptake by whole L. perenne was augmented by RSBC and CSBC. Accordingly, this work suggests that RSBC and CSBC have the potential to be used as a useful aided phytoremediation technology in Cd-contaminated soil.

Bioreduction of hexavalent chromium using a novel strain CRB-7 immobilized on multiple materials.

In this study, a novel Cr(VI) tolerant strain CRB-7 identified as Bacillus sp., was isolated and characterized for its high Cr(VI) reduction. The strain CRB-7 grew well and effectively reduced Cr(VI) under various conditions including pH (7-9), temperature (30-40 °C) and Cr(VI) concentrations (50-250 mg L-1). It almost completely reduced 120 mg L-1 Cr(VI) within 48 h under optimized condition of pH 7 and 37 °C. Further characterization by SEM-EDS and FTIR analyses indicated Cr(VI) removal mechanism of CRB-7 was predominately via bioreduction with little amount of bioadsorption. Furthermore, the strain CRB-7 based immobilized biobeads were successfully synthesized using five different porous materials as bacterial loading carrier respectively to ascertain the optimal immobilization biocomposite for Cr(VI) removal. CRB-7 cells immobilized with 3% sodium alginate (SA) and 5% humic acid (HA) exhibited the highest Cr(VI) removal efficiency. Moreover, immobilized biobeads have the advantages over free cells in being more stable and easier to reuse. High Cr(VI) reducing ability of the free and immobilized CRB-7 cells suggest the strain CRB-7, especially the B-HA-SA biocomposite is promising for remediating Cr(VI)-contaminated sites.