Chlorpyrifos (CPF) is one of the most widely used organophosphate pesticides worldwide. Epidemiological studies on pregnant women and their children suggest a link between in utero CPF exposure and delay in psychomotor and cognitive maturation. A large number of studies in animal models have shown adverse effects of CPF on developing brain and more recently on endocrine targets. Our aim was to determine if developmental exposure to CPF affects social responsiveness and associated molecular neuroendocrine markers at adulthood.Pregnant CD1 outbred mice were fed from gestational day 15 to lactation day 14 with either a CPF-added  or a standard diet. We then assessed in the offspring the long-term effects of CPF exposure on locomotion, social recognition performances and gene expression levels of selected neurondocrine markers in amygdala and hypothalamus.No sign of CPF systemic toxicity was detected. CPF induced behavioral alterations in adult offspring of both sexes: CPF-exposed males displayed enhanced investigative response to unfamiliar social stimuli, whereas CPF-exposed females showed a delayed onset of social investigation and lack of reaction to social novelty. In parallel, molecular effects of CPF were sex dimorphic: in males CPF increased expression of estrogen receptor beta in hypothalamus and decreased oxytocin expression in amygdala; CPF increased vasopressin 1a receptor expression in amygdala in both sexes.These data indicate that developmental CPF affects mouse social behavior and interferes with development of sex-dimorphic neuroendocrine pathways with potential disruptive effects on neuroendocrine axes homeostasis. The route of exposure selected in our study corresponds to relevant human exposure scenarios, our data thus supports the view that neuroendocrine effects, especially in susceptible time windows, should deserve more attention in risk assessment of OP insecticides. An increasing body of data collected in the past decade supports the developmental neurotoxicity of organophosphate insecticides (OPs), and of the most widely applied of them, chlorpyrifos (CPF), in particular. Most non-agricultural uses of CPF such as residential control of insect pests were phased out in the United States in 2001  and expord trimester) and CPF (2nd trimester) has been recently evidenced .All experiments on animals were performed according to the European Community Council Directive 86/609/EEC and to Italian Legislation on Animal Experimentation (Legislative Decree 116/92). Experiments were effectively carried out according to the Animal Research: Reporting In Vivo Experiments (ARRIVE) guidelines for reporting animal studies.Male and female mice of a Swiss-derived outbred strain , were housed in polycarbonate breeding cages with a 12–hr light-dark cycle (light on 20:00-8:00) and with free access to food and water. Females were inspected daily for the presence of the vaginal plug . The stud was removed 10 days after the discovery of the vaginal plug. On GD 15 females were randomly assigned to one of the two prenatal treatments , and it does not induce overt toxic symptoms in dams or major effects on pup’s health parameters such as weight at delivery and impairment of growing rate . All femg/kg/day .Sample size for behavioral tests was estimated to allow the detection of a standardized mean difference (delta effect size) = 1.2 between CPF and OIL groups, with a two-sided alpha = 0.05 and a power = 0.80. At these conditions, a minimum of n = 9 mice per group were needed. In the Open Field test, where male and female littermates were analyzed, at least 9 couples per treatment group were considered .Twenty litters (10 STD and 10 CPF) were left undisturbed till weaning, and at this time males and females from each litter were caged separately. Animals from each litter were assigned to experimental procedures following this scheme: one male and one female were assigned to behavior analysis and one male and one female from each litter were assigned to neuroendocrine assessment at adulthood. Slight differences in sample size between experiments were due to technical problem during video-recording.Fourteen (5 STD and 9 CPF) additional dams were assigned to the assessment of CPF effect on acetylcholinesterase (AChE) activity in brain and blood 24 h within delivery. AChE activity was further assessed on tissue samples on PND 0  and on PND 14  that were littermates of subjects undergoing behavioral and neuroendocrine assessments at adulthood.g at 4°C for 5 min) to separate plasma from red blood cells. Serum was collected and frozen for subsequent analysis.Mouse brains were removed from skull, weighed and homogenized (1 min) in cold 0.038 M Tris–HCl buffer, pH 7.6. Blood samples were collected centrifuged . A videocamera suspended over the arena was connected to a videotracking system  to record and analyze spontaneous behavior in the arena. The following variable were considered: distance moved in the arena to assess locomotor activity levels; time spent and number of visit in central and peripheral areas to assess preference for safer zone, and speed. Furthermore, we analyzed the time course of distance moved by dividing the 20 min total observation in four 5-min blocks.Males and females from each diet group underwent a social recognition test which was administered following two different protocols according to sex. As for males, the test consisted of repeated exposure to a con-specific kept under a small wire cage to prevent direct “aggressive” responses between the two males. As for females, we selected a direct free interaction protocol, since no aggressive responses occur during female-female mouse encounters. Direct free interaction protocols in female mice have greater resolution power since they allow to record not only behavioral responses but also ultrasonic vocalizations  -41. Impo19 male mice  were tested individually. The social recognition test took place on 2 days in a sound attenuated chamber. The social recognition task was preceded by a habituation session: each experimental subject was introduced for 10 min in the experimental cage enriched with a small round wire cage; after this period experimental mice underwent the social recognition test: Test (S1) - a stimulus mouse, unfamiliar con-specific of the same sex and age, was introduced for 5 minutes inside the home cage enclosed in the small, round wire cage; Retest same (S2) - after an inter-trial interval of 15 min the S1 stimulus mouse is re-introduced in the home cage enclosed in the wire cage for 5 min; Retest different (S3) - after an inter-trial interval of 15 min, a novel stimulus mouse was enclosed in the same wire cage and introduced in the home cage for 5 min. The entire social recognition test was videorecorded and later analyzed by an experimenter blind to treatment condition of the male “The Observer”, a software system for collection and analysis of behavioral data . During video-recorded sessions , the following behavioral responses were considered : social investigation, sniffing the round wire cage and the stimulus animal; rearing, standing on hind legs; grooming, self-cleaning .18 female mice  were individually housed in standard cages for 3 days and thus considered as resident females. Additional untreated females (n = 3) (of comparable age and body weight) socially housed served as test partners. The social recognition test took place on the following day in a sound attenuated chamber. Test (S1) - a female partner is introduced in the home cage of the isolated resident for 3 min and after that returned to its home cage; Retest Same (S2) - 45 min after the test, the same female partner is re-introduced in the home cage of the resident for 3 min; Retest Different (S3) - 45 min after the Retest Same, a novel partner is introduced for 3 min in the cage of the resident. For each resident, partners used in S2 and S3 phases were never drawn from the same social group to provide the maximum olfactory novelty during the S3 phase. During the three experimental phases number of ultrasonic vocalizations (USVs) was assessed using a ultrasonic microphone  sensitive to frequencies between 10-180 kHz and suspended 10 cm above the cage. Vocalizations were recorded using an Avisoft Recorder (Version 3.2). Settings included sampling rate at 250 kHz; format 16 bit. For acoustical analysis, recordings were transferred to Avisoft SASLab Pro (Version 4.40) and a fast Fourier transformation (FFT) was conducted. Spectrograms were generated with an FFT-length of 512 points and a time window overlap of 75% . The spectrogram was produced at a frequency resolution of 488 Hz and a time resolution of 1 ms. A lower cut-off frequency of 15 kHz was used to reduce background noise outside the relevant frequency band to 0 dB. Call detection was provided by an automatic threshold-based algorithm and a hold-time mechanism (hold time: 0.005 s). An experienced user checked the accuracy of call detection, and obtained a 100% concordance between automated and observational detection. The broad range for USV recording allowed detecting all USVs emitted but did not provide information about possible different meanings of such USVs (e.g. appetitive versus aversive meaning). During the test, the 3 min female–female encounters were also recorded by a videocamera placed 12 cm far from the cage. Videotapes were later analyzed by an experimenter blind to treatment condition of the resident females using “The Observer”, a software system for collection and analysis of behavioral data . During video-recorded sessions , frequency and duration of the following behavioral responses emitted by the resident female were collected: – exploring: moving around the cage, sniffing the physical environment, rearing; – social investigation: sniffing any part of the partner’s body; – grooming: self-cleaning .Oxt (Neurophysin I OXT carrier protein), Avp (Neurophysin II AVP carrier protein), OxtR, AvpR1A, ERα, ERβ, as well as for glyceraldehyde-3-phosphate dehydrogenase (Gapdh) and β-actin (Actb), as reference genes, were designed using the Primer-BLAST web application (www.ncbi.nlm.nih.gov/tools/primer-blast) and purchased from Invitrogen . Primers sequences are listed in Table Plus SYBR (Quantace) was used to perform Real-time PCR assays running reactions on a Stratagene MX5005P thermocycler instrument. Experiments were performed in duplicate on 96 well PCR plates. The thermal program was as follows: 1 cycle at 94 for 10 minutes; 40 cycles at 94°C for 10 seconds, 58°C for 10 seconds and 72°C for 10 seconds; 1 dissociation cycle from 55 to 94°C to verify amplification products. Evaluation of Gapdh and Actb stability was performed with BestKeeper  and not affected by CPF exposure. Only a main effect of sex was found [F  = 0.021 p < 0.01], indicating a higher level of locomotion in females  in each session of the test than STD mice [F  = 8.42 p = 0.001]. As expected in the third session of the social recognition test both CPF and STD showed a renewal of interest towards the unfamiliar mouse [F  = 12.78 p < 0.001]  = 5.93 p = 0.028] and a treatment x session interactions [F  = 3.79 p = 0.034] were observed. CPF females showed increased social investigation only during the second exposure to the same partner (S2)   expression, only a main effect of sex [Chi-Square (1) =8.652 p = 0.03] was detected.By contrast, Oxt expression in males . A CPF-induced increase was found for AvpR1a expression in both sexes   applied. However it is noteworthy that CPF males are affected throughout all the sessions of the social recognition test, an effect more evident during the first two session of the test. CPF enhanced social response also in females, but with a different pattern: enhanced response was evident only during the second exposure to the same partner , a peculiar profile confirmed by USV emission rates. These behavioral data confirm the results of the prenatal or neonatal exposure protocols where CPF treatment induced enhanced response towards social salient stimuli . ThuERβ expression in hypothalamus and decreased Oxt expression in amygdala; CPF also increased AvpR1a expression in amygdala in both sexes.As for neuroendocrine markers, both hypothalamus and amygdala appear as sensitive target areas for CPF exposure: as for males, CPF increased levels of ERβ are suggestive of mechanistic hypothesis to be further explored. The observation that CPF significantly induced ERβ expression in the hypothalamus and not in the amygdala suggests that CPF does not act as a general estrogen-like compound globally up-regulating ERβ. The selective CPF effect on ERβ in the absence of effect on ERα, could be explained by the existence of different ontogenetic expression patterns of ERα/ERβ transcripts [The sex- and region-specific effects of developmental CPF on nscripts ,46. Siminscripts .Oxt and AvpR1a expression in amygdala. This interference has to be interpreted together with behavioral results. Of note the enhanced investigative responses to unfamiliar social stimuli observed in CPF-exposed mice  is associated with a decrease of Oxt expression in amygdala in males. Intriguingly, the behavioral profile of mice carrying deletion of the Oxt gene [ERβ gene had no consequences on social recognition in either male or female mice [CPF also impacts on Oxt gene  also disale mice .Oxt and AvpR1a upregulation represents the focal CPF-induced change underlying behavioral alterations.All together, these data confirm our previous hypothesis that exposure to CPF leads to inappropriate processing of the salience of diverse social cues . Such hyin vivo study exploring potential adverse effects of CPF on molecular endpoints in amygdala. Future studies should focus on the links between the pattern of neuroendocrine effects observed in amygdala with selected downstream pathways known to be modulated by hypothalamic neuropeptides [This is the first peptides  and invopeptides ,25,53,54peptides ,56.in vitro methods. CPF alters gonadotropin-releasing hormone biosynthesis in GT1-7 hypothalamic cell lines [in vitro data from a wide screening study using cancer fibroblast cells (MCF-7BUS) evidenced that CPF selectively increases ERβ, but not ERα gene expression [ERβ is a target of CPF endocrine disrupting activity.A limited number of evidence is so far available exploring the endocrine disrupting potential of CPF, primarily exploring ll lines ; both anll lines  and estrll lines ,60, havepression . Our stuInterestingly, data from AChE in dams  show that opposite to a 80% reduction of serum AChE activity, no reduction of brain AChE activity was detected. These data partially confirm the order of magnitude of inhibition “ plasma > RBC > brain” developed by the PBPK/PD model developed for CPF in rat and humans, with inhibition of brain AChE occurring only at doses of 10 mg/Kg or higher . Other rThe route of exposure selected in our study corresponds to relevant human exposure scenarios since it assures continuous levels of exposure from pregnancy to lactation. It is worth noting that, at variance from previous findings showing small but significant brain AChE inhibition in the offspring 4 and 24 hrs following administration of CPF , we did in vitro, in animal models and in humans coherently supports that endocrine effects, especially in susceptible time windows, should deserve more attention in the risk assessment of OP insecticides for a twofold reason. On one side, CPF neuroendocrine effects on Oxt and AvpR1a are extremely relevant since these pathways are implicated in several affective and emotional behaviors extending to psychopathology [ERβ expression pattern could be associated with specific prenatal/perinatal morphogenetic pathways  whicnization .


The disparity in species richness among evolutionary lineages is one of the oldest and most intriguing issues in evolutionary biology. Although geographical factors have been traditionally thought to promote speciation, recent studies have underscored the importance of ecological interactions as one of the main drivers of diversification. Here, we test if differences in species richness of closely related lineages match predictions based on the concept of density-dependent diversification. As radiation progresses, ecological niche-space would become increasingly saturated, resulting in fewer opportunities for speciation. To assess this hypothesis, we tested whether reef fish niche shifts toward usage of low-quality food resources (i.e. relatively low energy/protein per unit mass), such as algae, detritus, sponges and corals are accompanied by rapid net diversification. Using available molecular information, we reconstructed phylogenies of four major reef fish clades  to estimate the timing of radiations of their subclades. We found that the evolution of species-rich clades was associated with a switch to low quality food in three of the four clades analyzed, which is consistent with a density-dependent model of diversification. We suggest that ecological opportunity may play an important role in understanding the diversification of reef-fish lineages. Explaining the wide differences in species richness among different evolutionary lineages has been a major goal of evolutionary biology since Darwin and still remains one of the greatest challenges of modern biology We used molecular phylogenies to test the hypothesis of diversity-dependent diversification in reef fishes, one of the most speciose vertebrate assemblages dating back at least to the early Tertiary (50 Ma) We focused on four representative clades with good phylogenetic resolution  reef fish families sensu: LabridaThe Pomacentridae (damselfishes and anemonefishes) is one of the four most speciose reef fish families with approximately 380 species The Chaetodontidae (butterflyfishes and bannerfishes) includes over 120 species with spectacular coloration and considerable ecological and morphological diversity The Acanthuroidei is a monophyletic suborder that includes the families Acanthuridae (surgeonfishes), Zanclidae (Moorish idol), and Luvaridae (Louvar) et al.et al.The taxon sampling and molecular data in this study were obtained from previous phylogenetic studies regarding the four abovementioned groups. DNA sequences were downloaded from Genbank following accession numbers in (1) Tang The sampled Chaetodontidae and Pomacentridae include all currently recognized genera . In the ClustalX 2.0 We inferred the phylogenetic relationships within each family through Bayesian Inference (BI) in MrBayes 3.1.2 using the GTR+Γ model http://cbsuapps.tc.cornell.edu/mrbayes.aspx), and results were analyzed with Tracer 1.5 To infer each tree using Bayesian Inference, two independent Markov Chain Monte Carlo (MCMC) runs (each with four chains), were implemented simultaneously for 10 million generations, with trees sampled and saved every 1,000 generations  using MrBayes Divergence times of lineages in all four groups of study were estimated using BEAST 1.5.4 et al.Since the vast majority of reported chaetodontid fossils are demonstrably erroneous For each of the four groups we used CBSU Web Computing Interface to run three independent analyses of 10 million generations, sampling trees every 500 generations. Resulting log files were examined using Tracer 1.5 While it is self-evident that all diets are nutritionally adequate, there are fundamental differences in food composition (i.e. C/N ratio per unit mass) We have classified all species according to their trophic modes and mapped them onto the BI trees using parsimony, as implemented in Mesquite 2.75 All diversification statistics were performed in R G) for each of the four groups using the method of Margallon & Sanderson , thus, we estimated rG under different values of ε . Subclades were defined at the highest monophyletic clades in which species share the same trophic mode. The species richness of such lineages was measured based on valid names listed in the Catalog of Fishes r and s, using a broken-stick distribution as the null expectation 2 distributed with 2 d.f.). Our hypothesis predicts that the flexible model would be more likely than the constant model especially when assigning higher net diversification rates at the origin of clades containing low-quality diet feeders. However, it does not necessarily mean that the net diversification rates have increased at that node. Alternatively, the clade could have retained an ancestral but elevated rate, while another clade or clades exhibited a decline in net diversification. To test these scenarios, we used a constrained version of the two-rate model (‘rate-decrease model’), where the highest net diversification rate must occur in the tree bipartition containing the root node, such that no net rate increase is allowed on the path from the root to the clade assigned by the shifting point We estimated global diversification rate  in the resulting statistics compiled with Tracer. All resulting phylogenies are shown in Mean evolutionary rate, Yule process birth rate, coefficients of variation and mean covariance are presented in Acanthurus + Ctenochaetus was 20 Ma (95% HPD: 31–10 Ma).In the Acanthuroidei, the 95% HPD interval was high for all ages at nodes corresponding to basal relationships. However, the family Acanthuridae had the origin estimated at 61 Ma (95% HPD: 86–50 Ma) . CharactChaetodon. The specialization of obligate corallivores appears shortly thereafter at about 15 Ma (95% HPD: 24–12 Ma), in the clade Chaetodon 3  was estimated at 55 Ma (95% HPD: 65–50 Ma) as we set in our prior. As expected, the family Chaetodontidae seems to be a relatively young family originated at 33 Ma (95% HPD: 45–21 Ma) . The reletodon 3 .For the labrid tree, three clades used for calibration had age estimates close to the age of the oldest fossil: labrid root  = 58 Ma (95% HPD: 62–55 Ma), Labridae 1 = 51 Ma (95% HPD: 53–50), and Labridae 2 = 54 Ma (95% HPD: 59–50) . CharactLepidozygus. There are three other clades with low-quality diets: Pomacentridae 4a , Pomacentridae 4b  and the genus Pomacentrus , from which only Pomacentrus seems to have arisen after the Oligo-Miocene transition (The origin of Pomacentridae estimated at 52 Ma (95% HPD: 61–50 Ma) was very close to the age of the oldest fossil of the family used to set a hard lower bound prior . The expansition .G) decreased with increasing extinction rates, as already noted in the method description. Significantly higher than expected species richness under rG conditioned to different ε values are found in Acanthurus + Ctenochaetus  has a mean age of 52 Ma, however, 75% of these species belong to the subclade formed by Acanthurus + Ctenochaetus, which originated about 20 Ma. Therefore, we believe that all influence that led to higher than expected number of extant species in Acanthuridae (-Naso) came exclusively from the effects of the Acanthurus + Ctenochaetus diversification. We applied the test considering Acanthurus + Ctenochaetus species richness and we obtained significant values of p under even higher extinction rates (ε = 0.0 to ε = 0.99). For Labridae 2 h (Labridae) and Pomacentrus (Pomacentridae) we obtained significant values of p under ε = 0.0 to ε = 0.9. The remaining clades showed significant p values under ε = 0.0 to ε = 0.5 (Chaetodon (Chaetodontidae), Pomacentrus (Pomacentridae), Labridae 1a and Labridae 2b (Labridae).Estimated global net diversification rates (r ε = 0.5 . From thAcanthurus + Ctenochaetus in Acanthuroidei (ε = 0.0 to ε = 0.5), Chaetodon in Chaetodontidae (ε = 0.0), Labridae 2 h in Labridae (ε = 0.0 to ε = 0.99) and Pomacentrus in Pomacentridae (ε = 0.0 to ε = 0.5) (The comparisons between a constant model and a flexible model showed that the flexible model provided the best explanation for elevated net diversification rate at clades ε = 0.5) .Acanthurus + Ctenochaetus in the Acanthuroidei  huroidei , Chaetoddontidae  and Pomaentridae . With exChaetodon, Pomacentrus and Acanthurus + Ctenochaetus to Oligo-Miocene tectonic events and the expansion of reefs in the IWP Prognathodes in Chaetodontidae, It is reasonable to attribute the high diversity found in Our results show remarkable parallels to recent studies of fossil surgeonfishes (Acanthuridae) and rabbitfishes (Siganidae) Scarus and Chlorurus, showed higher than expected net speciation rates. Several explanations for the relatively high rates of diversification in this subclade have been proposed, including the presence of an intermandibular joint Within the Labridae, the low-quality feeders of the subclade Labridae 2b, which encompasses It is hard to determine whether low-quality diets are taxonomically more diverse than high-quality diets, given that fishes often exploit complex resources rather than specific food items There is a central conceptual difference between the present study and that of Harmelin-Vivien Evolutionary delayed origins of low-energy diets seems to be a widespread pattern within marine communities and has already been described for major metazoan clades such as Crustacea, Mollusca and Echinodermata in different phylogenetic scales The patterns seen in the Acanthuroidei, Chaetodontidae and Labridae are all consistent with the suggestion that the use of low-quality food resources is a derived condition which has evolved independently at least once in each clade Chaetodon during the Miocene  evolution seems to agree quite well with Harmelin-Vivien's  Miocene . The recChaetodon spp. bite the substratum to ingest food, and are primarily distinguished from each other by the surface they bite. However, the dietary differentiation among chaetodontid lineages seems to transition evolutionarily from high to low-quality diets . p<0.05 indicates significant speciose subclades. NS  =  not significant. Subclade labeled in bold  =  subclades with species using low-quality diets.(DOCX)Click here for additional data file.Table S2Likelihoods and Akaike Information Criterion for constant and flexible models of diversification under different extinction rates. Likelihoods (LH) and Akaike Information Criterion (AIC) for constant and flexible models of diversification under different extinction rates (ε). r is the constant diversification rate estimated across all lineages. p<0.05 indicates significant higher likelihood estimated for the flexible model, in which a diversification rate r1 shift to r2 at the subclade indicated. NS indicates that the flexible model was not significant assuming shifts in any of the subclades. Rate decreasing scenario was not significant in any case, except in Pomacentridae over ε = 0.5, which was not considered due the equivocal phylogenetic position of Lepidozygus.(DOCX)Click here for additional data file.


There aIt is well known that extended application of ATP to activate the P2X receptors for tens of seconds or minutes induces a remarkable increase in membrane permeability to large molecules of up to 900 Daltons, a phenotype often referred to as formation of large pores. This was originally documented in immune cells about three decades ago; ATP permeablized cell membranes to nucleotides  being the cation in intracellular and extracellular solutions, respectively  293 cells  and ethidium (6.5 Å × 11 Å × 13Å) (Browne et al., The commonly used fluorescent dyes are, however, considerably larger in size than NMDGIn parallel with these efforts to understand P2X receptor-dependent large pore formation, studies have accumulated evidence to show the importance of such receptor functionality. For example, P2X7 receptor-dependent large pore formation has been identified as a crucial factor associated with disease conditions such as chronic pain (Sorge et al., LW, L-HJ and DL led the discussion; EC contributed to the discussion and generated the structural models. L-HJ wrote the manuscript, and all authors commented the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


This study investigates the biokinetics of LGT proteome, a potential biomarker of severe TBI, in serum of severe TBI patients. The LGT proteome presents in the serum of severe TBI patients. The abundance diversity of LGT proteome is closely associated with pathologic condition of TBI patients. Serum LGT proteome may be used as a promising marker for evaluating severity of severe TBI. Traumatic brain injury  is the most common disease with high morbidity and mortality in neurosurgery, especially to the youngsters, and a lot of researchers have focused on this fact , 2. SeveSerum protein biomarkers have long held promise in the study of TBI. They are considered to facilitate the diagnosis, monitoring progress, prognosis, and providing pertinent molecular information about ongoing pathological changes for designing evidence-based therapeutic interventions . Serum pBased on the findings above, LGT can also be found in severe TBI patients' serum, and it may be the early warning sign for disease aggravation or even death. In order to investigate the clinical significance of LGT proteome, the study proceeded with surface enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) , 13 techP > 0.05).96 TBI cases treated in our department between March 2006 and July 2009 were analyzed retrospectively. There were 77 males and 19 females with age range of 16–83 years (mean age: 31.7 ± 26.3); interval time from injury to clinical (h): 0.5~11.5 h, average 7.1 ± 2.2; Glasgow Coma Scale (GCS) score: 44 cases with 3~5 points, 52 cases with 6~8 points, and 32 cases receiving emergency operation; trauma cause: 46 cases by traffic accident, 22 cases by injury due to fall or mine disaster, 19 cases by strike injury, and 6 cases with other causes; major trauma type: 47 cases with contusion and laceration of brain and (or) subdural/intracerebral hematoma, 14 cases with epidural hematoma, and 35 cases with diffuse axonal injury and brain stem injury. All cases were diagnosed as craniocerebral trauma by cranial CT scanning after the admission; 69 survival cases carried on prognostic evaluation with Glasgow outcome score (GOS) 6 months later. The exclusion criteria were as follows: being under 16 years old, death in 24 hours from admission, severe infection or combined injury, malignant tumor or autoimmune disease, with damage of important organs especially liver and kidney (multiple organ failure), and transfusion of blood in operation. Meanwhile, 35 healthy cases were chosen as control group. This project has been reviewed by the Medical Ethics Committee of Shanxi Provincial People's Hospital. At the same time, signed informed consent of research projects of every patient with patients themselves or their guardians was obtained. The factors such as general condition, age, and sex were comparable  between the two groups ], HPLC water, and SPA.μL serum sample and add 10 μL urea (9 mmol/L). The sample underwent oscillation for 30 min, followed by the addition of 180 μL buffer , at 4°C, and was centrifuged at 10000 g × 2 min; then the supernatant fluid was taken for use.5 mL sera were taken in the morning when patients were hollow and were put in VACVTTE vacuum quantitative mining container and placed at room temperature for 60 min (including the time when samples were out of the body), and then the serum was separated from blood at 3000 g × 10 min and preserved at −80°C; serum samples were thawed and put in ice bath (10000 g × 2 min); take 5 μL combined cushioning liquid and carry out oscillation for 5 min at room temperature, discard liquid, and then repeat the procedure; then continue with each hole plus 100 μL serum sample at room temperature and carry out oscillation for 1 h. Discard the liquid and wash the hole with 200 μL combining buffer 2 times (5 min each time) and then with HPLC water. Dismantle the chip, perform natural drying, and add 0.5 μL SPA at each point, twice. Read the meter reading chip with WCX2 protein microarray after drying.Chip was put into the container with hydrochloric acid (100 mmol/L), oscillated for 4~5 min, and then washed with HPLC water; put the chip into the bioprocessor: start with each hole plus 200 M/Z) in the range 1.0 × 103~2.0 × 103, and collected position 20~80, with each point collected 20 times; the total points were 140 times. Before each collection, all-in-one protein microarray calibration instrument was used to reduce the error to less than 0.1%. Using the quality control protein array to do the repetitive detection, the peak and strength variation coefficient (SVC) are controlled below 0.05% and 15%, respectively.The same parameters were used for all the samples; data were collected using Ciphergen ProteinChip 3.2 analysis software. Set inspection sample laser intensity at 195, detection sensitivity at 8, collected data quality load ratios (M/Z) between 11100 + HDa and 11900 + HDa exhibit cluster sample of fingerprint which is LGT existing performance. Observe LGT proteome abundance values and the LGT dynamically in trauma group and control group.In the fingerprint, qualitative load ratios (x ± SD), and multiple comparisons of the average number of diversity samples were examined through SNK-q test; the count data was examined through χ2 test. SPSS11.0 was used for statistical analysis. P < 0.05 was considered statistically significant.All measurement data was represented with mean ± standard deviation , but there were no significant differences between groups ① and ②  subdural/intracerebral hematoma, and ③ epidural hematoma and/or skull fractures, analyzing three groups of LGT proteome abundance values. Results showed that the LGT proteome abundance values of groups ① and ② were significantly higher than of group ③, and the difference was statistically significant ( ① and ② .P < 0.05). With the progression of the course (days 7 and 14), there was no significant difference between the two groups (P > 0.05) , LGT proteome abundance value of death group was significantly higher than of the survival group ( > 0.05) .β) [α (TNF-α) [Traumatic brain injury (TBI) is the leading cause of death and disability among young adults . Severe β) , 22–25, β) , 26, gliβ) , 22, 27,β) , nerve gβ) , doublecβ) , high-moβ) , and so β) , 29, 30,β) , 32, gliβ) , Interleβ) , tumor n (TNF-α) , cell-fr (TNF-α) , and so Recently, Pei et al. –10 foundM/Z between 11100 + HDa and 11900 + HDa. One of the spectrum abundance values was ≧20%, but all abundance values were ≧5%, compared with the upstream and downstream peak cluster; the M/Z values were more than 1000 quality units, with striking background as well as outstanding fingerprint characteristics. So take the peak more than 5% for LGT proteome positive diagnosis standard [Identification and purification of candidate biomarkers is a critical step in the biomarker development process, since it provides insight into the disease biology and facilitates the development of analyte-specific assays . Technicstandard –10.M/Z; the upstream cluster peak got more than 1000 mass units (M/Z) than downstream cluster peak; it showed the same result as the finding of Pei [P < 0.01). The abundance and positive rate of LGT proteome of GCS 3–5 severe TBI group were significantly higher than of GCS 6–8 severe TBI group (P < 0.01). Decreased serum LGT level is associated with GCS scores. The results showed that the LGT proteome could be detected in serum of TBI patients; the abundance of LGT proteome is closely associated with pathogenesis of TBI patients. TBI often involves a combination of mechanical trauma and local hypoxemia, on which serum biomarker concentrations may provide critical therapeutic data to evaluate the brain injury. Serum biomarker after TBI could be used as a source for the identification of the brain injury, the extent of the injury, and the time of its occurrence and even for identifying its most likely outcome [The experiment proceeded through SELDI-TOF-MS technology; the LGT proteome was detected in the proteome fingerprint with 100% relative abundance. A fingerprint marker just like a cluster appeared between 11100 Da and 11900 Da g of Pei –10, so i outcome .P < 0.01), but it did not reveal a significant difference between group ① and group ②. The result showed that the LGT proteome had close relationship with the severity of craniocerebral trauma. Neuronal and glial cell loss results from necrotic and apoptotic cell death during the primary and secondary injury and may lead to an increase in various neuron- and glia-specific proteins in serum [Our present study divided the TBI patients into three groups: ① brain stem injury and diffuse axonal injury, ② contusion and laceration of brain and (or) subdural/intracerebral hematoma, and ③ epidural hematoma and (or) skull fracture. The result showed that the LGT proteome abundance in groups ① and  ② was significantly higher than in group ③ ; there was no significant difference between the two groups in days 7 and 14 (P > 0.05). So the LGT proteome abundance at the early pathogenetic stage could be used to predict the different prognosis of TBI patients; the higher the LGT proteome abundance was, the worse the prognosis was. The experiment also showed that the LGT proteome abundance of death and survival groups increased in day 7. It happened during the secondary craniocerebral injury period, so the increase of the LGT proteome abundance may be caused by the secondary craniocerebral injury. There is a second wave of long-lasting pathological changes called the secondary injury mechanism. These pathologic features include metabolic changes, neuroinflammation, vascular abnormalities, and neuronal and glial cell death [The experiment also analyzed the LGT proteome abundance during the dynamic change of severe TBI patients. The LGT proteome abundance was higher in death group than in survival group in the initial pathologic stage and then decreased in the two groups as the course was prolonged. At the early pathologic stage (days 1 and 4), the LGT proteome abundance was significantly higher in death group than in survival group . The LGT proteome presents in the serum of severe TBI patients. The abundance diversity of LGT proteome is intimately associated with pathogenetic condition of TBI patients. Serum LGT proteome may be used as a promising marker for evaluating severity of severe TBI.


T method relative to control animals. This data article presents complementary results related to the research article entitled ‘Acute restraint stress induces specific changes in nitric oxide production and inflammatory markers in the rat hippocampus and striatum’ This data article provides additional evidence on gene expression changes in the neuronal and inducible isoforms of nitric oxide synthase in the medial prefrontal cortex following acute stress. Male Wistar rats aged 6–8 weeks were exposed to control or restraint stress conditions for up to four hours in the dark cycle after which the brain was removed and the medial prefrontal cortex isolated by cryodissection. Following RNA extraction and cDNA synthesis, gene expression data were measured using quantitative real-time PCR. The mRNA levels of the neuronal and inducible nitric oxide synthase isoforms, and the inhibitory subunit of NF-κB, I kappa B alpha were determined using the ΔΔC Specifications table•The first genomic data demonstrating a single acute stress is capable of inducing increased neuronal NOS mRNA expression.•The first regional observation of simultaneous increases in neuronal and inducible NOS mRNA in the medial prefrontal cortex.•These data provide evidence for rapid inhibition of the inflammatory NF-κB pathway in the medial prefrontal cortex.We have presented genomic data from the medial prefrontal cortex in support of the research article entitled ‘Acute restraint stress induces specific changes in nitric oxide production and inflammatory markers in the rat hippocampus and striatum’ published in Free Radical Biology and Medicine Fig. 32.1Rattus norvegicus) aged 6–7 weeks postnatal were housed individually in a colony room and given ad libitum access to standard rat chow and water. This room was on a 12 h light–dark cycle (lights off at 12.30 h) and maintained under standard laboratory conditions . Procedures performed were approved by The University of Queensland Animal Ethics Committee with AEC approval number 018/11.Outbred male Wistar rats , 60, 120, and 240 minutes (stress treatment starting at 13:30 h) within individual home cages according to Spiers and colleagues 2.4n=5–6 per group). At the end of each treatment period, rats were sacrificed with sodium pentobarbital  and the brain removed for storage at −80 °C. Frozen brains were sectioned on a cryostat and the medial prefrontal cortex was cryo-dissected from sections according to a rat brain atlas Animals were randomly allocated to 0 (control), 60, 120, or 240 min stress groups . The mRNA expression was determined by real-time PCR using Taqman gene expression ‘assay-on-demandTM’ kits . The primer/probe sets analyzed were FAM-labeled Nos1 , Nos2 , and Nfkbia . Each primer/probe was analyzed in reactions multiplexed and normalized with a VIC-labeled primer/probe assay for GAPDH . The mRNA levels were normalized to the average of the control group using the ΔΔCT method.Total RNA was extracted from each regionalized medial prefrontal cortex using the RNeasy mini kit  and treated with deoxyribonuclease 1  according to the manufacturer׳s instructions. Extracted RNA was reverse transcribed into cDNA using the iScript2.6p-values less than 0.05 were considered statistically significant.Data were analyzed using statistical software GraphPad Prism . One-way repeated measure ANOVAs with Dunnet׳s multiple comparisons test were used to compare normally distributed data. A non-parametric Kruskal–Wallis ANOVA with Dunn׳s multiple comparisons test was used for data with significantly different standard deviations. Results were expressed as mean±standard error of the mean (±SEM) and


Under low light, the activity of the carbon-concentrating mechanism generally decreases, associated with an increase in leakiness (ϕ), the ratio of CO2 retrodiffusing from the bundle sheath relative to C4 carboxylation. This increase in ϕ had been theoretically associated with a decrease in biochemical operating efficiency  under low light and, because a proportion of canopy photosynthesis is carried out by shaded leaves, potential productivity losses at field scale. Maize plants were grown under light regimes representing the cycle that leaves undergo in the canopy, whereby younger leaves initially developed under high light and were then re-acclimated to low light  for 3 weeks. Following re-acclimation, leaves reduced rates of light-respiration and reached a status of lower ϕ, effectively optimizing the limited ATP resources available under low photosynthetically active radiation. Direct estimates of respiration in the light, and ATP production rate, allowed an empirical estimate of ATP production rate relative to gross assimilation to be derived. These values were compared to modelled ATP/GA which was predicted using leakiness as the sole proxy for ATP/GA, and, using a novel comprehensive biochemical model, showing that irrespective of whether leaves are acclimated to very low or high light intensity, the biochemical efficiency of the C4 cycle does not decrease at low photosynthetically active radiation.C C4 photronments . Under o3 plants  and very4 plants results from anatomical and biochemical differentiation of the leaf parenchyma. Externally mesophyll cells and internally bundle sheath (BS) cells are coupled to operate a biochemical carbon-concentrating mechanism (CCM) that increases the CO2 concentration in BS, the cellular compartment where Rubisco is exclusively expressed, resulting in active suppression of the oxygenase activity of Rubisco. Since BS and mesophyll cells are connected by plasmodesmata, some CO2 retrodiffuses (CO2 leakage). The extent of CO2 retrodiffusion is still debated, but it is accepted that the permeability to CO2 diffusion , defined as the rate of CO2 retrodiffusing (leak rate) relative to the phosphoenolpyruvate (PEP) carboxylation rate (VP). Since Rubisco CO2 fixation (in BS) is complementary to leakage (out of BS), ϕ can be used as a proxy for the coordination between the CCM and C3 assimilatory activity ; hence, anatomical features are tightly bound to biochemical and energy traits. Plants with a higher gBS would have higher rate of CO2 retrodiffusion, increased CCM cost, and a higher ATP demand for gross assimilation (ATP/GA), which is the overall biochemical operating efficiency of C4 photosynthesis. For this reason, ϕ has been used to derive ATP/GA . We suggested several mechanisms whereby C4 leaves adapted throughout growth to low-light conditions could maintain high photosynthetic conversion efficiency during steady-state photosynthesis.Previously we studied how long-term acclimation to low light influenced short-term responses to illumination . Plants JATP) was derived from gas exchange measurements under low O2 in combination with photosystem II (PSII) photochemical yield, measured CO2 assimilation rate, and online isotopic discrimination during photosynthesis (Δ). A full isotopic discrimination model was used to derive ϕ from Δ  could be calculated and compared with the predicted ATP cost of assimilation (ATP/GA). Mature leaves that had re-acclimated under low light (HLLL) showed very similar traits to LL plants. HLLL plants deployed two strategies to optimize the scarce ATP resources under low light: (i) the reduction of respiration in the light (RLIGHT) and (ii) the reduction of leakiness (ϕ). The comparison of JATP/GA with ATP/GA estimated with a novel metabolic model showed that C4 photosynthetic efficiency was constant in the vicinity of the light compensation point (LCP): thus, the predicted decrease in biochemical conversion efficiency based on ϕ increasing under limiting light does not occur.In this study we grew maize plants under a light regime representing the acclimation of leaves shaded by an over-growing canopy, consisting of 3 weeks under high light followed by 3 weeks under diffuse, low light intensity. The leaf-level ATP-production rate (ϕ from Δ . With th−2·s−1) and low-intensity diffuse light (PAR = 100 μE·m−2·s−1)—obtained using shading to mimic the understory of a canopy. Maize seeds  were sown weekly in 1.5 l pots filled with Levington pro M3 pot and bedding compost . Plants were grown in three sets of conditions: (i) HL plants were grown for 3 weeks under high light (fully expanded fourth leaf stage); (ii) LL plants were grown for 4 weeks under low light (fully expanded fourth leaf stage); and (iii) HLLL plants were grown for 3 weeks under high light, the youngest fully expanded leaf was marked, and then plants were grown for the following 3 weeks under low light. Plants were manually watered daily, with particular care to avoid overwatering. When ready, plants were measured once and then discarded. Measurements were performed on the youngest fully expanded leaf of HL and LL plants, and on marked leaves of HLLL plants.Plants were grown at the Plant Growth Facility located at the University of Cambridge Botanic Garden in controlled-environment growth rooms  set at 16-h day length, 25/23 °C (day/night), and 40% relative humidity. The growth protocol was designed to standardize age and watering conditions throughout the experiment. Two light environments were established—high-intensity direct light  and either a mixture of 2% O2/N2 or ambient air. Photosystem I (PSI) yield and PSII yield (Y(II); see −2·s−1, enough to saturate fluorescence and PSI signals . The block temperature was set at 26 °C so as to maintain the leaf temperature close to 25 °C. The IRGA was connected to a cryogenic H2O- and CO2-trapping purification line. Each day, one plant was subject to a RGB-light-response curve, under 2% O2 and Ca = 600 μmol·mol−1 [to determine the relationship between electron transport rate (ETR) and JATP] and a second RGB-light-response curve under 21% O2 and reference CO2 set at 400 μmol·mol−1, during which exhaust gas was trapped to determine Δ. With this procedure each day the δ13C composition of a total of 12 CO2 samples and six CO2 references  were analysed directly using a VG SIRA dual-inlet isotope ratio mass spectrometer . Δ was calculated as reported in Y(II) was determined at each light level for both light curves. JATP was calculated individually at each irradiance by multiplying the relationship between ETR and JATP (determined at low O2) by the ratio between Y(II) at ambient and low O2 3 (−1) or with values of RLIGHT estimated through non-linear curve fitting of light-response curves 3 . Althougϕ was resolved from Δ using the full model of Farquhar, as recently integrated to take into account the ‘ternary’ effects, i.e. the effect of water molecules diffusing outward stomata on CO2 molecules diffusing inwards through still air (4 photosynthesis are summed. This model requires the CO2 concentration in the different cellular compartments (notably mesophyll and BS cells), which were calculated by means of the validated C4 photosynthesis model, in the light-limited form  and RLIGHT. BS conductance, required to parameterize the C4 model, cannot be measured directly but it can be estimated by fitting the C4 model to a measured quantity. In the ‘Δ/Δ’ fitting  was calculated from JATP and net assimilation, A. The derivation of JATP through the low O2-ETR method was described above  was calculated using values for GA, derived by summing A plus RLIGHT calculated by curve fitting (see above). JATP/GA is relevant to C4 biochemistry and shows the empirical conversion efficiency of CO2 into sugars. It is worth stressing that these values for JATP/GA are derived with a novel method based on gas exchange under low O2  is generally dependent on light intensity and hence it is only marginally influenced by O2 partial pressure  while the ATP cost of the CCM depends solely on ϕ. This implies that the CCM is driven solely by the activity of PEPC and that all the retrodiffusing CO2 is refixed. Under these assumptions the ATP cost of the CCM is calculated by multiplying the overall ATP cost of PEPC (2 ATP/CO2) by the ratio of CO2 overcycling [1/(1−ϕ)]. The total ATP cost of gross assimilation results from summing the cost of the C3 activity plus the cost of the CCM  and light intensities from 50 to 500 μE·m−2·s−1 using the values of ϕ derived from isotopic discrimination.Here the subscript GA, whereby the ATP demand of all biochemical processes underpinning assimilation (hence B approach) are summed. The B approach is comprehensive, and requires the quantification of all processes contributing to C4 photosynthesis. We used the validated C4 model  rate, and PPDK is the pyruvate phosphate dikinase (PPDK) rate. PEPCK was assumed to regenerate 20% of the PEP required by PEPC, which is close to the expected value under natural white light (VP), VC, and VO were calculated with the validated von Caemmerer C4 model . For the purposes of these calculations, rather than using a fixed value of x, such as 0.4  significantly differentiated plant responses , but as light decreased between PAR 250 and 0 μE·m−2·s−1 the response approached that of LL plants. Consistently, the LCP and RLIGHT of HLLL plants were similar to those of LL plants, and clearly lower than those of HL plants (esponses . LL planL plants .JATP) is shown in JATP was derived from gross assimilation under low O2 and then corrected for photorespiration at ambient O2 using the ratio of photochemical yield. At high PAR, JATP tracks the pattern of A; however, at low PAR, JATP of all plants was similar, suggesting that the higher A of LL and HLLL plants at limiting PAR (inset in Δ) is shown in Δ increased substantially at PAR lower than 250 μE·m−2·s−1, although in HLLL plants Δ was, on average, lower than for HL plants. LL plants showed a more gradual increase under decreasing PAR.The total ATP production rate (inset in  was achiinset in . Isotopigs) and Ci/Ca. Ci/Ca differentiated clearly between growth conditions, and was lowest in LL plants and highest in HL plants, while HLLL plants had intermediate values at all levels of PAR. Ci/Ca was higher than 0.5 at PAR <125 μE·m−2·s−1 (LL plants), reflecting the efforts made during the measurements to induce stomatal opening. A high Ci/Ca was functional in the resolution of the isotopic discrimination model, to maximize the contribution of biochemical processes over the stomatal contribution to total isotopic discrimination , which was estimated by fitting a C4 photosynthesis model, rearranged to express JMOD, to the values for JATP described above. The difference between conditions was not significant, and was due to a small difference in permeability to CO2 retrodiffusion out of the BS . HL plants had higher ϕ than LL and HLLL under limiting PAR, with ϕ increasing from 0.25 to 0.35 under decreasing PAR. HLLL plants had the lowest ϕ under light intensities higher than 75 μE·m−2·s−1. Under low light intensities they showed an increase in ϕ with a similar trend to that of HL plants. In LL plants ϕ was close to 0.24 and only marginally affected by light intensity.on model , paramet fitting . ϕ signiJATP/A. This quantity expresses the ATP cost involved in the assimilation of CO2; that is, how much ATP the plant has to produce  to assimilate one CO2 molecule. JATP/A for HLLL plants was very similar to that of LL plants and significantly lower than that of HL plants. This means that re-acclimation was extremely effective in reducing JATP/A, particularly in the vicinity of the LCP. JATP/GA. This quantity is the biochemical conversion efficiency of C4 assimilation, or how much ATP is needed to convert bicarbonate into stable assimilates. The empirical values for JATP/GA  assimilation were derived. GA with two different approaches. These methods are compared in ϕ as a sole proxy for C4 operating efficiency . A second strategy involved the reduction of leakiness (ϕ). HLLL plants showed reduced values for ϕ as compared to HL plants. However the ϕ hyperbolic increase under low irradiance was similar to that of HL plants. In contrast, and in agreement with recent results  was considered, only minor energetic differences could be observed between different light treatments. Re-acclimation significantly influenced neither the empirical JATP/GA  nor the predicted ATP cost of GA . This shows that if there were any effect of varied ϕ on the overall biochemical conversion efficiency the effect was undetectable using the methods described. On one hand this confirms the difficulties in estimating leakiness from leaf-level energetics  regulating reducing power export from mesophyll to BS cells in response to demand, (iii) partitioning metabolic work between contrasting cells types , (iv) optimizing energy availability in BS and mesophyll cells while at the same time (v) maintaining the equilibrium between the CCM and the C3 activity, and, finally, (vi) trading-off at the level of BS conductance, between the capacity to support very high diffusion (and assimilation) rates and the necessity to limit leakage of CO2 out of the BS  regu4 biochemical operating efficiency  uses leakiness, ϕ as the sole proxy . With the ϕ approach the C3 activity is considered to have an invariable cost of 3 ATP/GA (photorespiration is neglected), whereas the CCM is assumed to be supplied solely by PEPC activity, which is assumed to refix entirely the retrodiffused (leaked) CO2. Our empirical evidence largely confirms the validity of the ϕ approach, which closely predicted the trend and the magnitude of JATP/GA under PAR >125 μE·m−2·s−1 for LL + HLLL plants, and under PAR ≥500 μE·m−2·s−1 for HL plants. However, under low irradiances, the ϕ approach overestimated the trend of JATP/GA, especially for HL plants. This overestimation is dependent on the assumptions of the ϕ approach (photorespiration is neglected and the CCM is driven solely by PEPC), which hold only under high irradiances, while under low irradiance they are no longer valid. In fact, PEPC and Rubisco activities proportionally decrease under decreasing irradiance, limited by the decreasing ATP availability. As opposed to that, BS respiration is largely unaffected by light intensity, and, under decreasing light intensities, the BS-respired CO2 progressively outweighs PEP carboxylation rate (VP). Hence, ϕ—that is, the ratio of retrodiffusing CO2 over PEP carboxylation rate—becomes progressively higher as light intensity approaches the compensation point. This gives rise to the extensively documented  hyperbolic ϕ increase  and the optimal partitioning factor between the CCM activity and the C3 activity (x) are largely independent of light intensity , whereby the ATP cost of all processes contributing to assimilation are summed. We used the equations that we have recently derived  of the CCM under high light regimes. However, we showed that  4 model , 2013. Wnet assimilation under low light intensities, which, for HLLL plants, decreased by 35% as compared to HL plants under PAR = 50 μE·m−2·s−1. This shows clearly that the net energy conversion efficiency under limiting light is to a considerable extent ameliorated by the acclimation of mature leaves to low light.In this study we set out to investigate the strategies deployed by maize plants grown under high light intensities when re-acclimated to low light. We showed that the main re-acclimation drivers were the reduction of respiration and the reduction of leakiness, and these were likely to be accompanied by complex metabolic reorganization. Overall, these strategies were very effective in reducing the ATP cost of gross assimilation we could isolate the contribution of day respiration from the other biochemical effects (which include the reduction of leakiness). The ATP cost of gross assimilation was not significantly different for HLLL plants as compared to HL plants. This showed that re-acclimation did not change the efficiency of C4 metabolism, even if it considerably reduced leakiness, implying that the effect of reduced leakiness on C4 energetics was not detectable. Leakiness dynamics may then be associated to other processes occurring at biochemical level such as the regulation between BS versus mesophyll metabolic engagement and CCM versus C3 activity. In addition, we provided compelling theoretical and empirical evidence showing that the increase in hyperbolic leakiness, observed under low light intensities (4 efficiency loss under low light conditions (e.g. 4 model (4 conversion efficiency (as ATP cost of gross assimilation), taking into account the active and passive contributions to the CCM.By calculating ATP cost of ensities , is not ons e.g. . We propons e.g.  based on 4 model . The bioThe implications for loss of productivity at the field scale being specifically associated with increased leakiness (JXB online.Supplementary material is available at Supplementary Fig. S1. ξ values for the calculation of Δ (equations are reported in


Proton transfer across single-layer graphene proceeds with large computed energy barriers and is therefore thought to be unfavourable at room temperature unless nanoscale holes or dopants are introduced, or a potential bias is applied. Here we subject single-layer graphene supported on fused silica to cycles of high and low pH, and show that protons transfer reversibly from the aqueous phase through the graphene to the other side where they undergo acid–base chemistry with the silica hydroxyl groups. After ruling out diffusion through macroscopic pinholes, the protons are found to transfer through rare, naturally occurring atomic defects. Computer simulations reveal low energy barriers of 0.61–0.75 eV for aqueous proton transfer across hydroxyl-terminated atomic defects that participate in a Grotthuss-type relay, while pyrylium-like ether terminations shut down proton exchange. Unfavourable energy barriers to helium and hydrogen transfer indicate the process is selective for aqueous protons. Proton transfer across graphene is associated with large computed energy barriers and is thought to be generally unfavourable. Here, the authors observe aqueous proton transfer through graphene subjected to pH cycling, suggesting that it is due to transfer through rare, naturally occurring atomic defects. As shown in 141519a,eff values of 3.5(1) and 8.3(2), which fall within the reported literature values  images of graphene single layers deposited on fused silica windows show a low density of macroscopic pinholes and that the graphene is free of cracks or folds . Two-dims−1 refs , 38, 39.− groups . This re−5 cm2 s−1, or just half of that of bulk water, once protons arriving through any opening within the graphene sheet interact with the hydroxylated portion of the surface that is located behind the reaction front. To conservatively assess an upper bound limit for our estimations, we calculated the proton mean-square displacement using a D-value of 1 × 10−6 cm2 s−1. The probability of placing our laser beam within the propagating reaction front emanating from a given macroscopic pinhole was then estimated to be 4% and 21% for 1 and 10 s SHG jump times, respectively  was then used to search for atomic defects using annular dark field (ADF) STEM imaging at 60 kV (see Methods). The majority of the images show perfect six-fold symmetry in the position of the carbon atoms and vast areas that lack grain boundaries and atomic, or vacancy, defects . Neverth44To elucidate the mechanisms for proton transfer, we discuss findings from density functional theory (DFT) calculations  and Reax50We find that the main restriction for aqueous proton transfer through pristine, defect-free graphene is the energy required to push the proton through the center of an aromatic ring in the hydrophobic graphene layer as shown in The basal planes of pristine graphene can, and do, contain rare atomic-scale defect sites comprised of carbon atom vacancies, as shown in et al.)All of the defect terminations considered are energetically favourable as compared with the bare quad-vacancy system . The rem2 transfer requires barriers exceeding 1.9 eV  to fill ~1 cm2 with a single layer. Following annealing in a flow of 4% H2 in Ar for 30 min at 300 °C, vibrational sum-frequency generation spectra showed no evidence for CH stretchesWe used graphene having a grain size of ~100 μm ref.  grown on54SEM images were collected from the center of the graphene film. Graphene on an optical window was imaged using a Hitachi S-4800 scanning electron microscope (SEM) operating at 2 kV with a probing current of 10 μA and an Everhart–Thornley detector. Copper tape was used to reduce charging effects. Individual images were taken at × 1,200 magnification with 1,280 × 960 resolution. An array of 5 × 5 images  was stitched together using Adobe Illustrator. Automatic brightness and contrast adjustment on each frame was carried out using the ‘auto adjust’ feature in Preview . No other postedit feature or change was applied.−5 torr) for 8 h to remove surface contamination. Following heating in vacuum, the specimen was immediately transferred to the UltraSTEM for ADF STEM imaging. The surface of the graphene still retains residual PMMA that was used in the transfer processes to the transmission electron microscopy grid as shown in To confirm the single-layer nature of graphene synthesized using the CVD method2 and NaCl . The concentration of NaCl was confirmed using a conductivity meter . Solution pH was adjusted with minimum amounts of dilute solutions of ~1 M NaOH  and HCl (EMD ACS grade). The pH-jump experiments were carried out using a fused silica hemisphere  pressed against either a fused silica window  or a CVD-prepared graphene film transferred onto a silica window in an experimental setup previously reported13The aqueous solutions were prepared with Millipore water, prepared the day before an experiment and left open to air overnight to equilibrate with atmospheric CO−1 flow using variable flow peristaltic pumps as previously reported13As shown in 134 μJ cm−2 per pulse with a 30 μm focal spot, which is well below the damage threshold of graphene as previously reported1313A detailed description of our SHG setup has been described previously606162Ab Initio Simulation Package64x and y dimensions. A 15 Å gap was inserted between the graphene layer perpendicular to the surface. The gap was subsequently filled with enough water molecules to match the overall density of water at 1.0 × 103 kg m−3. The initial simulations were carried out with water on both sides of the graphene layer. The lower SiO2 substrate was initially simplified by using additional water. Subsequent calculations were carried out with more realistic slabs comprised of water/graphene/water/SiO2 substrates. The reaction rates and mechanisms of proton transfer through the graphene were described in the framework of transition state theory and within the harmonic approximation, which is robust for systems of high densities.First-principles periodic DFT calculations were carried out to determine the lowest energy interfacial water/graphene, water/graphene/water/silica structures and the activation barriers for proton diffusion through these interfaces using the Vienna −5 eV. All of the atoms were allowed to relax in the geometry optimizations until the forces on each atom were <0.03 eV Å−1. Spin polarization was examined for all of the systems explored and applied when needed. Transition states were isolated using the nudged elastic band method44All of the calculations were carried out within the generalized gradient approximation using Perdew–Burke–Ernzerhof functionalab initio MD to validate predictions of force field in describing water/graphene systems .Supplementary Figures 1-26, Supplementary Tables 1-3, Supplementary Notes 1-13 and Supplementary References


Symbiodinium. Elevated temperatures can result in the expulsion of Symbiodinium or loss of their photosynthetic pigments and is known as coral bleaching. It has been postulated that the expression of light-harvesting protein complexes (LHCs), which bind chlorophylls (chl) and carotenoids, are important in photobleaching. This study explored the effect a sixteen-day thermal stress (increasing daily from 25–34 °C) on integral LHC (chlorophyll a-chlorophyll c2-peridinin protein complex (acpPC)) gene expression in Symbiodinium within the coral Acropora aspera. Thermal stress leads to a decrease in Symbiodinium photosynthetic efficiency by day eight, while symbiont density was significantly lower on day sixteen. Over this time period, the gene expression of five SymbiodiniumacpPC genes was quantified. Three acpPC genes exhibited up-regulated expression when corals were exposed to temperatures above 31.5 °C . In contrast, the expression of acpPCSym_5:1 and acpPCSym_10:1 was unchanged throughout the experiment. Interestingly, the three acpPC genes with increased expression cluster together in a phylogenetic analysis of light-harvesting complexes.Coral reef success is largely dependent on the symbiosis between coral hosts and dinoflagellate symbionts belonging to the genus  Symbiodinium form symbiotic relationships with a variety of marine taxa. Endosymbiotic associations observed between these photosynthetic dinoflagellates and corals are generally classed as mutualistic, as both the host and symbiont benefit from the relationship1Symbiodinium are critical for the proliferation of reefs supporting diverse marine ecosystems. Coral bleaching, the loss of photosynthetic pigments or endosymbionts from host cells occurs under stress conditions such as elevated sea temperatures of only a few degrees above long-term maxima46Photosynthetic eukaryotic dinoflagellates belonging to the genus Symbiodinium and the coral holobiont has focussed on many environmental factors implicated in the onset of coral bleaching including elevated seawater temperatures, eutrophication and disease. The effect of high sea-surface temperatures have been a key focus due to mass coral bleaching events  complexes, which are found in dinoflagellates2122LHCs, also called antenna proteins, are found in photoautotrophic organisms and are an array of protein, chl and accessory pigment molecules with roles in light-harvesting and photoprotection17Symbiodinium two types of peripheral LHCs are found, PCP and acpPCa/c subfamily of LHCsSymbiodinium C3 acpPCs and Symbiodinium A1.1 LHCs cluster sequences with three clades within the chl a/c binding LHC family, indicating high diversity of these proteins within speciesSymbiodinium PCPs are well studied due to their unique features252627Symbiodinium the LHCs have been shown to decrease energy transfer and dissociate from the photosystem reaction centres following photoinhibition in order to protect cells during stress events12293031Symbiodinium acpPC expression16In Symbiodinium transcript levels have shown that changes in gene expression occur on a relatively small scale. Quantitative-PCR has been used to determine changes in a variety of genes of interest related to stress response333435Symbiodinium has allowed for a reference to be established in order to determine differential gene expression under various conditions36Symbiodinium physiology and large fold changes in host gene expression have been recorded in cells exposed to stress, changes in Symbiodinium gene expression occur at a far smaller scale (±<5-fold)3334Targeted studies of acpPC genes in Symbiodinium cells during a prolonged thermal stress event (sixteen days) in Acropora aspera. We find that the response of five acpPC genes in Symbiodinium under stress varies across a LHC phylogeny. This is the first study to investigate the effect of thermal stress on gene expression patterns in the LHC superfamily within a coral host.This study focussed on the response of A. aspera were exposed to temperature increasing from ambient levels (~25 °C) to a bleaching temperature of ~34 °C (Symbiodinium cell densities (p < 0.001) over the course of the experiment, with average densities of 6.0 × 105 cells cm−2 in the treatment corals on day sixteen, compared to 1.6 × 106 cells cm−2 in the control corals on the same day  and treatment  but not in the interaction treatment × day   and treatment  but not in the interaction treatment × day   . Similart period c. Analys df = 5) c. The raperiment d.Fv/Fm) was measured following sunset during the experiment. For corals maintained at control temperatures, Fv/Fm was between 0.602 and 0.692 (average 0.657)  and differences between day  and treatment   (Maximum quantum yield of photosynthesis (e 0.657) . Analysi df = 1) . A seque < 0.01) .Fv/Fm, there were significant interaction effects between treatment × day , and between day  and treatment   was also measured over the course of the experiment. As with  df = 1) . Post hoperiment .acpPC genes  from three distinct LHC clades , acpPCSym_15 on day twelve  and acpPCSym_18 on days ten  and sixteen   which both belong to Clade 3b  . These t= 0.020) , both acClade 3b  no signiClade 3b ,c. In adre found .acpPC genes in Symbiodinium under prolonged thermal stress of A. aspera and is the first to examine the expression of acpPC genes from different LHC Clades. The sixteen-day thermal regime was selected to enable sampling at temperatures leading up to, and inclusive of, a bleaching event and is the first experiment to investigate differential expression of integral antenna proteins in Symbiodinium within a coral host under thermal stress. Quantitative PCR was used to quantify the expression of five acpPC genes that are dispersed though out three clades of the chl a/c lineage of the LHC phylogeny  fluorometry analysis demonstrated that ficiency , this isd twelve  illustraacpPC genes was found to vary in Symbiodinium cells throughout the experiment. Functionally little is known about the diversity of LHCs, for example whether complexes only associate with specific photosystems, are some more efficient at light capture or energy transfer, are others favoured for photoprotection or do some display increased stability under high temperatures. Characterisation of SymbiodiniumacpPCs has shown that there is large diversity within the gene super-familySymbiodinium genome has provided more of an insight into the diversification of the LHC familySymbiodinium genome. The complexity observed in the integral LHC family has been attributed to multiple rounds of intra- and inter-genic gene duplication events202144acpPC may have different functions. As such, the differences in expression between those acpPC  are four pigment - protein complexes , and associated with PSII are six pigment – protein complexes Symbiodinium where there is high sequence diversity coupled with high copy number, and as yet it is not clear which proteins bind to PSI or PSII21Some ways in which psaA and psbA have previously been investigated in Symbiodinium under thermal stress. Decreases of psaA and psbA are hypothesised to significantly impair the mechanisms associated with coping with thermal stresspsbA gene, which encodes the core PSII D1 protein, was also quantified  on day ten of the thermal stress experiment. In Symbiodinium in hospite, these small changes in transcripts have been observed previously3334353347As in previous studies, investigating transcript abundance in acpPCSym_1:1, acpPCSym_15 and acpPCSym_18) were found to have increased expression over the duration of the experiment and interestingly, grouped in Clade 1 and Clade 2 of the LHC phylogeny. Additionally two LHC genes (acpPCSym_5:1 and acpPCSym_10:1) grouped with Clade 3b did not exhibit differences in expression. Though transcriptional changes were detected, expression changes observed were less than 2.5 fold throughout the experiment. This is consistent with previous studies where small-scale changes in gene expression were also observed. Given that we currently do not know how the diverse range of LHCs are associated with the photosystems, both PSII and PSI, their specific functional roles, , or the importance of Symbiodinium photosynthesis to the survival of corals, it is imperative that future research focuses on the specific roles of LHCs.This study exploited a bleaching experiment to investigate the effect of thermal stress on photosynthetic genes. Quantitative PCR was used to determine the expression of five integral LHC genes. Three LHC genes (n = 312) were collected (under Great Barrier Reef Marine Park Authority permit G13/36402.1) from four different colonies of Acropora aspera  on the reef flat of Heron Island at low tide in May 2013. Heron Island colonies of A. aspera have been demonstrated to associate only with Symbiodinium clade C33549Coral fragments  and thermal-stress treatment conditions. The control conditions remained at the ambient seawater temperature (~24 °C) for the duration of the experiment . FluctuaSymbiodinium within A. aspera. Imaging-PAM analysis was performed on the first day and every day from the third day following sunset. Nubbins were dark-adapted for twenty minutes prior to imaging-PAM analysis. Three replicate nubbins from each of the eight replicate tanks were designated for Imaging-PAM analysis and used throughout the experiment. Corals were measured in the same order at each imaging-PAM analysis. Dark-adapted yield and maximal fluorescence were determined using a weak pulse of light, followed by a saturating pulse of 2,700 μmol quanta m−2 s−1 of photosynthetically active radiation (PAR) for 800 ms. Induction recovery curves were used to examine the photosynthetic efficiency and ability of symbionts to recover from light stress throughout the experiment. The induction recovery analysis utilised fifteen pulses of saturating light  with a constant actinic light (111 μmol quanta m−2 s−1) over a 5-minute period, followed by a dark recovery period of 14 min. Data from the induction recovery curve was used to determine photo-kinetic parameters, such as Fv/Fm and NPQ. Following analysis, nubbins were returned to experimental tanks.Imaging-PAM fluorometry  was used to measure photosynthetic efficiency of n = 9–12) and were stripped of tissue using a Waterpik™ dental irrigator using seawater. On day zero the blastate was centrifuged at 3,076 g for 5 min immediately following tissue removal. On days eight, ten, twelve and sixteen the blastate was homogenised with an immersion blender for 5 s and centrifuged at 3,076 g for 3 min to pellet algal cells. Total pelleted cells were resuspended in 50 mL of seawater. Aliquots (1 mL) were taken for cell density approximation. The remaining cells were centrifuged at 3,076 g for 3 min to pellet cells and stored at −80 °C for chl a and c quantification. Chl was extracted in 90% acetone for 20 h in the dark at 4 °C and quantified using the equations of Jeffrey and Humphrey (1975)n = 5). Surface area of waterpiked nubbins was determined using the wax dipping methodAt 2 pm on days zero, eight, ten, twelve and sixteen, three replicate nubbins were taken from each replicate tank . Reverse transcription was performed using the SuperScript™ III First-Strand Synthesis SuperMix for qRT-PCR (Invitrogen). The 2x RT Reaction Mix contains the following components oligo (dT)20 (2.5 μM), random hexamers (2.5 ng.μl−1), 10 mM MgCl2, and dNTPs. Template cDNA dilution series were prepared to optimize quantification accuracy. For analysis, cDNA was diluted 1:40 prior to use as a template in the qRT-PCR analysis. Quantitative RT-PCR was performed using a Rotor-Gene™ 6000 . The qRT-PCR was performed in a final volume of 15 μl, containing 7.5 μl of GoTaq® qPCR Master Mix , 4 μl of diluted template and gene specific primers (266 nM). qRT-PCR conditions were as follows: 95 °C for 2 min; 40 cycles of 95 °C for 15 s and 61 °C for 60 s. All qRT-PCR were followed by a melt curve analysis from 55 to 95 °C to ensure single product amplification  and snap-frozen in liquid nitrogen and stored at −80 °C for later mRNA isolation. Coral branches that had been snap-frozen in liquid nitrogen were crushed with a hydraulic press before transfer to a mortar chilled with liquid nitrogen and ground finely with a chilled pestle. The powder was then divided into cryotubes and stored at −80 °C. Messenger RNA was isolated from cells using the Dynabeads® mRNA DIRECT™ kit as per the protocol outlined in Leggat fication . Each Rocyclophin (Cyc)glyceraldehyde 3-phosphate dehydrogenase (GAPDH)S4 ribosomal protein (Rp-S4)S-adenosyl-L-methionine synthetase (SAM)acpPC  Symbiodinium C3 primers were designed against acpPC sequences from an EST librarywww.ncbi.nlm.nih.gov). Primers were designed for acpPC genes in Symbiodinium using the software DNASTAR Primer Select (Lasergene 11) (psbA primers were previously established for use in Symbiodiniumhttp://www.biogazelle.com/products/qbasePLUS)5454Housekeeping genes used in our analysis were selected from previously established reference genes, which included Proliferating Cell Nuclear Antigen (PCNA)gene 11) . The psba and c, imaging-PAM and gene expression data. The sequential Bonferroni post hoc test was used to adjust for the false discovery rate (or type I error).Statistics software package  was used for all statistical analyses. A generalized linear model with ‘day’ and ‘treatment’ as main effects and ‘day × treatment’ as an interaction was used for pairwise comparisons of cell density, chl How to cite this article: Gierz, S. L. et al. Integral Light-Harvesting Complex Expression In Symbiodinium Within The Coral Acropora aspera Under Thermal Stress. Sci. Rep.6, 25081; doi: 10.1038/srep25081 (2016).


Experiments indicate that the ground state of graphene in a strong magnetic field exhibits spontaneous breaking of SU(4) symmetry. However, the nature of the corresponding emergent state is unclear because existing theoretical methods approximate the broken-symmetry solutions, yielding nearly-degenerate candidate ground states having different emergent orders. Resolving this ambiguity in the nature of the strong-field ground state is highly desirable, given the importance of graphene for both fundamental physics and technical applications. We have discovered a new SO(8) symmetry that recovers standard graphene SU(4) quantum Hall physics, but predicts two new broken-SU(4) phases and new properties for potential ground states. Our solutions are analytical; thus we capture the essential physics of spontaneously-broken SU(4) states in a powerful yet solvable model useful both in correlating existing data and in suggesting new experiments. Bc Bc on sample impurities suggests that the resistance is an intrinsic property of an emergent state differing qualitatively from perturbed SU(4) solutions (spontaneous symmetry breaking)Graphene in a strong magnetic field has approximate SU4) symmetry123 symmetry567analytically. As a first application we revisit the nature of the ground state for undoped monolayer graphene in a magnetic field.Here we show that SU(4) symmetry can be extended to an SO(8) symmetry that recovers graphene SU(4) physics, but that implies new low-energy modes that transcend SU(4) symmetry and for which solutions may be obtained sublattice pseudospin. The dispersion computed in tight-binding approximation11K and K′. The two-fold K degree of freedom is termed valley isospin. Near these K-points the dispersion is linear, leading to Dirac cones. For undoped graphene the Fermi surface lies at the apex of the cones, where the level density vanishes and the effective electronic mass tends to zero. Hence, near the K points low-energy electrons for undoped graphene in zero magnetic field obey a massless Dirac equation and behave mathematically as massless chiral fermions, with chirality related to projection of the sublattice pseudospin.Good reviews of graphene physics are available811n = 0 level is unusual in that it is half filled in the ground state of undoped graphene, leading to the anomalous counting observed in the graphene quantum Hall effect13K or K′), inter-valley tunneling may be ignored and the electrons in the valley reside entirely on either the A or B sublattice, implying that for the n = 0 LL valley isospin and sublattice pseudospin are equivalent labels. We shall be concerned primarily with this n = 0 Landau level, which has, in addition to the Landau orbital degeneracy, a 4-fold degeneracy corresponding to spin and valley isospin.In a magnetic field the massless Dirac equation may be solved with an appropriate vector potential and the resulting Landau levels (LLs) are labeled by integers. The n = 0 LL. Justification and caveats for this approximation are discussed in ref. 56The largest energy scales are the LL separation and Coulomb energy. For neutral graphene the LL separation is approximately three times larger than the Coulomb energy, which is in turn much larger than other interactions. Hence, we shall ignore inter-LL excitations and consider only a single τα operate on valley isospin, the Pauli matrices σα operate on electronic spin, gz and g⊥ are coupling constants, μB is the Bohr magneton, and the spin z direction is assumed aligned with the magnetic field. The three terms in where the Pauli matrices σ and the projection of the valley isospin τ are displayed in The four internal states representing possible combinations of the projection of the spin c†(c) create (annihilate) fermions, τ and σ are defined in mk labels orbitally-degenerate LL states. Physically, Tα is total valley isospin, Nα is a Néel vector measuring the difference in spins on the A and B sublattices, and the Παβ couple spin and valley isospin. Under commutation the operators (2–5) close an SU(4) algebra that commutes with the Coulomb interactiongz and g⊥. Four symmetry-breaking patterns have been discussedwhere rators 2– close anrators 2– close anN dimensional fermionic space the most general bilinear products N) Lie algebra under commutation. Adding the most general pair operators cicj extends SU(2N) to SO(4N)18N) will then represent the most general Hamiltonian for the collective subspace, and will be diagonal in the subspace basis for each dynamical symmetry chain. Thus, the manybody problem can be solved exactly in the symmetry limits defined by each subgroup chain2122232420262728For a 2-n = 0 LL with creation operators k is the LL orbital degeneracy, B is magnetic field, and S sample size. The fractional occupation of the LL is n is electron number, and the filling factor is For graphene we assume a single S† and Now we add to the 15 SU4) generators of  generatoa =  level and one in the b =  level, with the total mk of each pair coupled to zero term by term:where We also introduce for later use the linear combinationsThe 28 operators n = 0 LL using pairs similar physically to ours: FM order,CDW order, and AF order. Thus pairs in 5analytical solutions for corresponding collective modes.Pair configurations created by generators of Broken-symmetry states based on graphene SU(4) have been expressed in terms of the pair wavefunction5mk and the sum is over spin and valley labels. Now let us consider SO(8) pairs. All states of an irreducible representation may be constructed by successive application of raising and lowering operators to a highest-weight (HW) state (Cartan–Dynkin method)u denote the number of broken pairs. For u = 0 states at half filling the pair number is a = 1 and one in the a = 2 basis states  F(G) is specified by the Cartan–Dynkin procedure. For example, applying the isospin lowering operator Other states can be constructed by applying successively to u = 0 representation can be constructed by using successive applications of raising and lowering operators fashioned from the generators of Likewise, all other states of the exact manybody solution using standard techniques. We shall deal with these exact solutions in future papers. Here, we interpret the states implied by Eg depends only on n and a single order parameter β. For the symmetry groups gThe energy where the group-dependent coefficients are tabulated in ref. Cg denotes the quadratic Casimir invariant of g, and b2 and G0 are coupling strengths.where β distinguishes the phases associated with the subgroup chains in β measures AF, since it is related to the AF order parameter The order parameter Coherent states violate translational, rotational, and gauge invariance. However, for realistic fields and sample sizes these violations are negligible, yielding a Ginzburg–Landau type theory with microscopic pedigree.Closed forms are given for the SO(8) CS wavefunctions in ref. Q± pairs, each with vanishing S pairs, each with vanishing Thus the SU(4) state is a superposition of f = n/2Ω in β at the minimum, which is zero for f = 0.5 curves. These are shown in β.Energies for the q ~ 1, sinceQuantum phase transitions between symmetry limits may be studied by varying coupling. We rewrite G0 ~ b2. Variation of ground state energy with q is shown in q phase transitions may be initiated by changing particle occupancy. β = 0, through a critical SO(7) symmetry with energy highly degenerate in β, to SU(4) with β ≠ 0, as f is changed at constant q.implies SO(7) symmetry if single parameter β that measures AF order: SU(4) states have finite β and AF order, but no CDW or FM order, β = 0 and no AF, CDW, or FM order, and SO(7) states define a critical dynamical symmetry that interpolates between SU(4) and n = 0 LLThus SO(8) describes analytically a host of broken-SU(4) candidates for the states in graphene being unraveled in modern experiments73536Transport properties are not manifest in the algebraic theory but the CS approximation is equivalent to symmetry-constrained Hartree–Fock–Boboliubov (HFB) theory25G0 and b2 in Solutions depend on Tc superconductors (SC)28critical dynamical symmetry generalizing a quantum critical point to a quantum critical phase linking other phases through quantum fluctuations. These similarities may have implications for cross-disciplinary understanding of quantum phase transitions.The present ideas are similar to ones found in nuclear physicsTc superconductors, suggesting a deep mathematical connection among these phenomena.We have introduced an SO(8) model of monolayer graphene in a magnetic field that recovers SU(4) quantum Hall physics but implies new collective modes transcending explicitly-broken SU(4) that are leading candidates for the high-field ground state. Graphene SO(8) is isomorphic to a symmetry describing many complex nuclei and very similar to one describing high-How to cite this article: Wu, L.-A. and Guidry, M. The Ground State of Monolayer Graphene in a Strong Magnetic Field. Sci. Rep.6, 22423; doi: 10.1038/srep22423 (2016).